805 results on '"biotherapeutics"'
Search Results
2. Plant-derived nanoparticles and plant virus nanoparticles: Bioactivity, health management, and delivery potential.
- Author
-
Liu, Cun, Yu, Yang, Fang, Liguang, Wang, Jia, Sun, Chunjie, Li, Huayao, Zhuang, Jing, and Sun, Changgang
- Subjects
- *
PLANT viruses , *DRUG delivery systems , *CELL communication , *CHARACTERISTIC functions , *PATHOGENIC microorganisms - Abstract
Natural plants have acquired an increasing attention in biomedical research. Recent studies have revealed that plant-derived nanoparticles (PDNPs), which are nano-sized membrane vesicles released by plants, are one of the important material bases for the health promotion of natural plants. A great deal of research in this field has focused on nanoparticles derived from fresh vegetables and fruits. Generally, PDNPs contain lipids, proteins, nucleic acids, and other active small molecules and exhibit unique biological regulatory activity and editability. Specifically, they have emerged as important mediators of intercellular communication, and thus, are potentially suitable for therapeutic purposes. In this review, PDNPs were extensively explored; by evaluating them systematically starting from the origin and isolation, toward their characteristics, including morphological compositions, biological functions, and delivery potentials, as well as distinguishing them from plant-derived exosomes and highlighting the limitations of the current research. Meanwhile, we elucidated the variations in PDNPs infected by pathogenic microorganisms and emphasized on the biological functions and characteristics of plant virus nanoparticles. After clarifying these problems, it is beneficial to further research on PDNPs in the future and develop their clinical application value. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
3. Internalization of therapeutic antibodies into dendritic cells as a risk factor for immunogenicity.
- Author
-
Siegel, Michel, Bolender, Anna-Lena, Ducret, Axel, Fraidling, Johannes, Hartman, Katharina, Looney, Cary M., Rohr, Olivier, Hickling, Timothy P., Kettenberger, Hubert, Lechmann, Martin, Marban-Doran, Céline, and Kraft, Thomas E.
- Subjects
PEPTIDES ,DENDRITIC cells ,IMMUNE response ,SURFACE charges ,T cells - Abstract
Introduction: Immunogenicity, the unwanted immune response triggered by therapeutic antibodies, poses significant challenges in biotherapeutic development. This response can lead to the production of anti-drug antibodies, potentially compromising the efficacy and safety of treatments. The internalization of therapeutic antibodies into dendritic cells (DCs) is a critical factor influencing immunogenicity. Using monoclonal antibodies, with differences in non-specific cellular uptake, as tools to explore the impact on the overall risk of immunogenicity, this study explores how internalization influences peptide presentation and subsequently T cell activation. Materials and methods: To investigate the impact of antibody internalization on immunogenicity, untargeted toolantibodies with engineered positive or negative charge patches were utilized. Immature monocyte-derived DCs (moDCs), known for their physiologically relevant high endocytic activity, were employed for internalization assays, while mature moDCs were used for MHC-II associated peptide proteomics (MAPPs) assays. In addition to the lysosomal accumulation and peptide presentation, subsequent CD4+ T cell activation has been assessed. Consequently, a known CD4+ T cell epitope from ovalbumin was inserted into the tool antibodies to evaluate T cell activation on a single, shared epitope. Results: Antibodies with positive charge patches exhibited higher rates of lysosomal accumulation and epitope presentation compared to those with negative charge patches or neutral surface charge. Furthermore, a direct correlation between internalization rate and presentation on MHC-II molecules could be established. To explore the link between internalization, peptide presentation and CD4+ T cell activation, tool antibodies containing the same OVA epitope were used. Previous observations were not altered by the insertion of the OVA epitope and ultimately, an enhanced CD4+ T cell response correlated with increased internalization in DCs and peptide presentation. Discussion: These findings demonstrate that the biophysical properties of therapeutic antibodies, particularly surface charge, play a crucial role in their internalization into DCs. Antibodies internalized faster and processed by DCs, are also more prone to be presented on their surface leading to a higher risk of triggering an immune response. These insights underscore the importance of considering antibody surface charge and other properties that enhance cellular accumulation during the preclinical development of biotherapeutics to mitigate immunogenicity risks. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
4. Development and characterization of dendritic cell internalization and activation assays contributing to the immunogenicity risk evaluation of biotherapeutics.
- Author
-
Siegel, Michel, Padamsey, Aman, Bolender, Anna-Lena, Hargreaves, Patrick, Fraidling, Johannes, Ducret, Axel, Hartman, Katharina, Looney, Cary M., Bertinetti-Lapatki, Cristina, Rohr, Olivier, Hickling, Timothy P., Kraft, Thomas E., and Marban-Doran, Céline
- Subjects
MONONUCLEAR leukocytes ,IMMUNE response ,DENDRITIC cells ,DRUG bioavailability ,THERAPEUTIC use of proteins - Abstract
Introduction: Immunogenicity refers to the ability of a substance, such as a therapeutic drug, to elicit an immune response. While beneficial in vaccine development, undesirable immunogenicity can compromise the safety and efficacy of therapeutic proteins by inducing anti-drug antibodies (ADAs). These ADAs can reduce drug bioavailability and alter pharmacokinetics, necessitating comprehensive immunogenicity risk assessments starting at early stages of drug development. Given the complexity of immunogenicity, an integrated approach is essential, as no single assay can universally recapitulate the immune response leading to the formation of anti-drug antibodies. Methods: To better understand the Dendritic Cell (DC) contribution to immunogenicity, we developed two flow cytometry-based assays: the DC internalization assay and the DC activation assay. Monocyte-derived dendritic cells (moDCs) were generated from peripheral blood mononuclear cells (PBMCs) and differentiated over a five-day period. The internalization assay measured the accumulation rate of therapeutic antibodies within moDCs, while the activation assay assessed the expression of DC activationmarkers such as CD40, CD80, CD86, CD83, and DC-SIGN (CD209). To characterize these two assays further, we used a set of marketed therapeutic antibodies. Results: The study highlights that moDCs differentiated for 5 days from freshly isolated monocytes were more prone to respond to external stimuli. The internalization assay has been shown to be highly sensitive to the molecule tested, allowing the use of only 4 donors to detect small but significant differences. We also demonstrated that therapeutic antibodies were efficiently taken up by moDCs, with a strong correlation with their peptide presentation on MHC-II. On the other hand, by monitoring DC activation through a limited set of activation markers including CD40, CD83, and DC-SIGN, the DC activation assay has the potential to compare a series of compounds. These two assays provide a more comprehensive understanding of DC function in the context of immunogenicity, highlighting the importance of both internalization and activation processes in ADA development. Discussion: The DC internalization and activation assays described here address key gaps in existing immunogenicity assessment methods by providing specific and reliable measures of DC function. The assays enhance our ability to pre-clinically evaluate the immunogenic potential of biotherapeutics, thereby improving their safety and efficacy. Future work should focus on further validating these assays and integrating them into a holistic immunogenicity risk assessment framework. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
5. Revolutionizing disease treatment through bioengineered probiotics and glucagon‐like peptide 1 (GLP‐1) based strategies: A path towards effective cures.
- Author
-
Jain, Smriti, Shukla, Adarsh Kumar, Deepika, Panwar, Surbhi, Kumari, Anita, Yadav, Ashok Kumar, and Kumar, Ashwani
- Subjects
GUT microbiome ,JUNK food ,PROBIOTICS ,PEPTIDES ,DIETARY patterns - Abstract
Human intestinal gut microbiota harbors complex and diverse microbes that play an important role in maintaining the homeostasis of the intestinal microenvironment in humans. The rise in mortality and morbidity rates among humans because of the increased incidence of food‐borne pathogens and the habits of individuals to eat junk food poses greater concerns and needs to be addressed. Bioengineering of probiotics has enabled the researchers to advance their research by developing probiotics with more functionalities. Moreover, GLP‐1 peptides which are incretin hormones have been shown to be more effective when combined with engineered probiotics. Various studies have shown its effectiveness in diabetic mice where human‐modified GLP‐1 produced long‐lasting benefits and research is going on to study its role in other diseases. The role of designer probiotics in treating and preventing diseases have been of much interest in recent times. However, the role of GLP‐1 peptides in treating diseases and their efficacy in combination with next‐gen biotherapeutics have received little attention. Thus, this review enlightens about the baseline knowledge as well as knowledge gaps related to conventional and genetically engineered probiotics. It also discusses the effect of GLP‐1 peptides in combination with bioengineered probiotics to prevent and treat diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
6. Pulmonary Inhalation of Biotherapeutics: A Systematic Approach to Understanding the Effects of Atomisation Gas Flow Rate on Particle Physiochemical Properties and Retained Bioactivity.
- Author
-
Foley, Laura, Ziaee, Ahmad, Walker, Gavin, and O'Reilly, Emmet
- Subjects
- *
GAS flow , *GRANULAR flow , *ENGINEERS , *LYSOZYMES , *BIOPHARMACEUTICS - Abstract
The identification of spray-drying processing parameters capable of producing particles suitable for pulmonary inhalation with retained bioactivity underpins the development of inhalable biotherapeutics. Effective delivery of biopharmaceuticals via pulmonary delivery routes such as dry powder inhalation (DPI) requires developing techniques that engineer particles to well-defined target profiles while simultaneously minimising protein denaturation. This study examines the simultaneous effects of atomisation gas flow rate on particle properties and retained bioactivity for the model biopharmaceutical lysozyme. The results show that optimising the interplay between atomisation gas flow rate and excipient concentration enables the production of free-flowing powder with retained bioactivity approaching 100%, moisture content below 4%, and D50 < 4 µm, at yields exceeding 50%. The developed methodologies inform the future design of protein-specific spray-drying parameters for inhalable biotherapeutics. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
7. Limosilactobacillus fermentum Strains as Novel Probiotic Candidates to Promote Host Health Benefits and Development of Biotherapeutics: A Comprehensive Review.
- Author
-
de Luna Freire, Micaelle Oliveira, Cruz Neto, José Patrocínio Ribeiro, de Albuquerque Lemos, Deborah Emanuelle, de Albuquerque, Thatyane Mariano Rodrigues, Garcia, Estefânia Fernandes, de Souza, Evandro Leite, and de Brito Alves, José Luiz
- Abstract
Fruits and their processing by-products are sources of potentially probiotic strains. Limosilactobacillus (L.) fermentum strains isolated from fruit processing by-products have shown probiotic-related properties. This review presents and discusses the results of the available studies that evaluated the probiotic properties of L. fermentum in promoting host health benefits, their application by the food industry, and the development of biotherapeutics. The results showed that administration of L. fermentum for 4 to 8 weeks promoted host health benefits in rats, including the modulation of gut microbiota, improvement of metabolic parameters, and antihypertensive, antioxidant, and anti-inflammatory effects. The results also showed the relevance of L. fermentum strains for application in the food industry and for the formulation of novel biotherapeutics, especially nutraceuticals. This review provides evidence that L. fermentum strains isolated from fruit processing by-products have great potential for promoting host health and indicate the need for a translational approach to confirm their effects in humans using randomized, double-blind, placebo-controlled trials. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
8. Immunogenicity of Atezolizumab: Influence of Testing Method and Sampling Frequency on Reported Anti-drug Antibody Incidence Rates.
- Author
-
Usdin, Maxime, Quarmby, Valerie, Zanghi, James, Bernaards, Coen, Liao, Laura, Laxamana, Joel, Wu, Benjamin, Swanson, Steven, Song, Yuan, and Siguenza, Patty
- Abstract
Measurement of anti-drug antibodies (ADA) to assess the incidence of ADA in a clinical trial is a critical step in immunogenicity assessment during the development of a protein therapeutic. We developed novel graphical approaches to illustrate clinical trial ADA data for the PD-L1 inhibitor atezolizumab (Tecentriq) that included a systematic analysis of the impact of the timing of ADA sampling and ADA assay drug tolerance on reported ADA incidence. We found that approaches used across the industry for ADA incidence analysis provide a limited view of immunogenicity in oncology studies, where ADA detection may be confounded by both drug dosage and patient attrition. Moreover, these approaches can miss important temporal information about the immune response. Our results demonstrated that the methodology of ADA assessment for the atezolizumab program was specifically designed to capture most ADA responses to ensure accurate reporting of ADA incidence. We further showed that the use of sparse sampling and/or ADA test methods with insufficient drug tolerance may result in a significant underreporting of ADA incidence. We conclude that the comparison of ADA incidence between different drugs can be highly misleading and that a test method with appropriate sensitivity in the presence of the drug and a clinical sampling scheme that is aligned with ADA responses to a drug is required to accurately report ADA incidence. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
9. Revolutionizing disease treatment through bioengineered probiotics and glucagon‐like peptide 1 (GLP‐1) based strategies: A path towards effective cures
- Author
-
Smriti Jain, Adarsh Kumar Shukla, Deepika, Surbhi Panwar, Anita Kumari, Ashok Kumar Yadav, and Ashwani Kumar
- Subjects
bioengineered probiotics ,biotherapeutics ,GLP‐1 ,GLP‐1 receptors ,probiotics ,Food processing and manufacture ,TP368-456 - Abstract
Abstract Human intestinal gut microbiota harbors complex and diverse microbes that play an important role in maintaining the homeostasis of the intestinal microenvironment in humans. The rise in mortality and morbidity rates among humans because of the increased incidence of food‐borne pathogens and the habits of individuals to eat junk food poses greater concerns and needs to be addressed. Bioengineering of probiotics has enabled the researchers to advance their research by developing probiotics with more functionalities. Moreover, GLP‐1 peptides which are incretin hormones have been shown to be more effective when combined with engineered probiotics. Various studies have shown its effectiveness in diabetic mice where human‐modified GLP‐1 produced long‐lasting benefits and research is going on to study its role in other diseases. The role of designer probiotics in treating and preventing diseases have been of much interest in recent times. However, the role of GLP‐1 peptides in treating diseases and their efficacy in combination with next‐gen biotherapeutics have received little attention. Thus, this review enlightens about the baseline knowledge as well as knowledge gaps related to conventional and genetically engineered probiotics. It also discusses the effect of GLP‐1 peptides in combination with bioengineered probiotics to prevent and treat diseases.
- Published
- 2024
- Full Text
- View/download PDF
10. Secretome from iPSC-derived MSCs exerts proangiogenic and immunosuppressive effects to alleviate radiation-induced vascular endothelial cell damage
- Author
-
Kshama Gupta, Ralph B. Perkerson, Tammee M. Parsons, Ramacharan Angom, Danilyn Amerna, Jeremy D. Burgess, Yingxue Ren, Pamela J. McLean, Debabrata Mukhopadhyay, Prasanna Vibhute, Zbigniew K. Wszolek, Abba C. Zubair, Alfredo Quiñones-Hinojosa, and Takahisa Kanekiyo
- Subjects
iPSC-MSC ,Radiation therapy ,Angiogenesis ,Inflammation ,Secretome ,Biotherapeutics ,Medicine (General) ,R5-920 ,Biochemistry ,QD415-436 - Abstract
Abstract Background Radiation therapy is the standard of care for central nervous system tumours. Despite the success of radiation therapy in reducing tumour mass, irradiation (IR)-induced vasculopathies and neuroinflammation contribute to late-delayed complications, neurodegeneration, and premature ageing in long-term cancer survivors. Mesenchymal stromal cells (MSCs) are adult stem cells that facilitate tissue integrity, homeostasis, and repair. Here, we investigated the potential of the iPSC-derived MSC (iMSC) secretome in immunomodulation and vasculature repair in response to radiation injury utilizing human cell lines. Methods We generated iPSC-derived iMSC lines and evaluated the potential of their conditioned media (iMSC CM) to treat IR-induced injuries in human monocytes (THP1) and brain vascular endothelial cells (hCMEC/D3). We further assessed factors in the iMSC secretome, their modulation, and the molecular pathways they elicit. Results Increasing doses of IR disturbed endothelial tube and spheroid formation in hCMEC/D3. When IR-injured hCMEC/D3 (IR ≤ 5 Gy) were treated with iMSC CM, endothelial cell viability, adherence, spheroid compactness, and proangiogenic sprout formation were significantly ameliorated, and IR-induced ROS levels were reduced. iMSC CM augmented tube formation in cocultures of hCMEC/D3 and iMSCs. Consistently, iMSC CM facilitated angiogenesis in a zebrafish model in vivo. Furthermore, iMSC CM suppressed IR-induced NFκB activation, TNF-α release, and ROS production in THP1 cells. Additionally, iMSC CM diminished NF-kB activation in THP1 cells cocultured with irradiated hCMEC/D3, iMSCs, or HMC3 microglial lines. The cytokine array revealed that iMSC CM contains the proangiogenic and immunosuppressive factors MCP1/CCL2, IL6, IL8/CXCL8, ANG (Angiogenin), GROα/CXCL1, and RANTES/CCL5. Common promoter regulatory elements were enriched in TF-binding motifs such as androgen receptor (ANDR) and GATA2. hCMEC/D3 phosphokinome profiling revealed increased expression of pro-survival factors, the PI3K/AKT/mTOR modulator PRAS40 and β-catenin in response to CM. The transcriptome analysis revealed increased expression of GATA2 in iMSCs and the enrichment of pathways involved in RNA metabolism, translation, mitochondrial respiration, DNA damage repair, and neurodevelopment. Conclusions The iMSC secretome is a comodulated composite of proangiogenic and immunosuppressive factors that has the potential to alleviate radiation-induced vascular endothelial cell damage and immune activation.
- Published
- 2024
- Full Text
- View/download PDF
11. Secretome from iPSC-derived MSCs exerts proangiogenic and immunosuppressive effects to alleviate radiation-induced vascular endothelial cell damage.
- Author
-
Gupta, Kshama, Perkerson III, Ralph B., Parsons, Tammee M., Angom, Ramacharan, Amerna, Danilyn, Burgess, Jeremy D., Ren, Yingxue, McLean, Pamela J., Mukhopadhyay, Debabrata, Vibhute, Prasanna, Wszolek, Zbigniew K., Zubair, Abba C., Quiñones-Hinojosa, Alfredo, and Kanekiyo, Takahisa
- Subjects
- *
VASCULAR endothelial cells , *ANDROGEN receptors , *GENE expression , *STEM cells , *PREMATURE aging (Medicine) , *RNA metabolism , *DNA repair - Abstract
Background: Radiation therapy is the standard of care for central nervous system tumours. Despite the success of radiation therapy in reducing tumour mass, irradiation (IR)-induced vasculopathies and neuroinflammation contribute to late-delayed complications, neurodegeneration, and premature ageing in long-term cancer survivors. Mesenchymal stromal cells (MSCs) are adult stem cells that facilitate tissue integrity, homeostasis, and repair. Here, we investigated the potential of the iPSC-derived MSC (iMSC) secretome in immunomodulation and vasculature repair in response to radiation injury utilizing human cell lines. Methods: We generated iPSC-derived iMSC lines and evaluated the potential of their conditioned media (iMSC CM) to treat IR-induced injuries in human monocytes (THP1) and brain vascular endothelial cells (hCMEC/D3). We further assessed factors in the iMSC secretome, their modulation, and the molecular pathways they elicit. Results: Increasing doses of IR disturbed endothelial tube and spheroid formation in hCMEC/D3. When IR-injured hCMEC/D3 (IR ≤ 5 Gy) were treated with iMSC CM, endothelial cell viability, adherence, spheroid compactness, and proangiogenic sprout formation were significantly ameliorated, and IR-induced ROS levels were reduced. iMSC CM augmented tube formation in cocultures of hCMEC/D3 and iMSCs. Consistently, iMSC CM facilitated angiogenesis in a zebrafish model in vivo. Furthermore, iMSC CM suppressed IR-induced NFκB activation, TNF-α release, and ROS production in THP1 cells. Additionally, iMSC CM diminished NF-kB activation in THP1 cells cocultured with irradiated hCMEC/D3, iMSCs, or HMC3 microglial lines. The cytokine array revealed that iMSC CM contains the proangiogenic and immunosuppressive factors MCP1/CCL2, IL6, IL8/CXCL8, ANG (Angiogenin), GROα/CXCL1, and RANTES/CCL5. Common promoter regulatory elements were enriched in TF-binding motifs such as androgen receptor (ANDR) and GATA2. hCMEC/D3 phosphokinome profiling revealed increased expression of pro-survival factors, the PI3K/AKT/mTOR modulator PRAS40 and β-catenin in response to CM. The transcriptome analysis revealed increased expression of GATA2 in iMSCs and the enrichment of pathways involved in RNA metabolism, translation, mitochondrial respiration, DNA damage repair, and neurodevelopment. Conclusions: The iMSC secretome is a comodulated composite of proangiogenic and immunosuppressive factors that has the potential to alleviate radiation-induced vascular endothelial cell damage and immune activation. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
12. Optimization, physicochemical stability and in vivo study of alginate-chitosan composites as nanocarriers for low molecular weight angiotensin I-converting enzyme (ACE)-inhibitory peptide.
- Author
-
Shehu Muhammad Auwal, Siti Balqis Muhammad Ghanisma, and Nazamid Saari
- Subjects
- *
BIOTHERAPY , *ALGINATES , *IN vitro studies , *T-test (Statistics) , *DATA analysis , *HYPERTENSION , *ACE inhibitors , *FISHER exact test , *ANTIHYPERTENSIVE agents , *IN vivo studies , *NANOMEDICINE , *DRUG delivery systems , *DESCRIPTIVE statistics , *POLYSACCHARIDES , *PEPTIDES , *BIOMEDICAL materials , *RATS , *FOOD industry , *CALCIUM chloride , *DRUG efficacy , *ANIMAL experimentation , *ANALYSIS of variance , *STATISTICS , *BIOSIMILARS , *DATA analysis software , *NANOPARTICLES , *ANGIOTENSIN I , *ANALYTICAL chemistry , *PHARMACODYNAMICS , *EVALUATION - Abstract
Chitosan and alginate, are non-toxic and biodegradable polymers used to enhance the stability of biotherapeutics by loading them into nanocarriers. In this study, the stone fish-derived low molecular weight peptide (Ala-Leu-Gly-Pro-Gln-Phe-Tyr), exhibited an in vitro ACE-inhibitory activity of 94.43 ± 2.05% and an IC50 of 0.012 ± 0.001 mM. The peptide was encapsulated via ionic gelation with alginate followed by polyelectrolyte complexation with chitosan. The resulting ACE-inhibitory peptide-loaded alginate-chitosan nanoparticles (ACE-I-ALG-CS NPs) were optimized to achieve small particle size (212.60 nm) and high encapsulation efficiency (EE, 74.48%). This was based on an optimum chitosan concentration (0.420%w/v), homogenization speed (6000 rpm), and homogenization time (30 min) using Box Behnken experimental design (BBED). Characterization of the ACE-I-ALG-CS NPs revealed a spherical, monodispersed morphology with high physicochemical stability during storage at 2 °C, 7 °C, and 12 °C for 12 weeks. Moreover, the in vivo study conducted on spontaneously hypertensive rats (SHRs) demonstrated a significantly higher (p < 0.05) systolic blood pressure (SBP)-lowering effect of the ACE-I-ALG-CS NPs compared to captopril and unencapsulated peptide. Hence, alginate and chitosan can be used as biocompatible coating materials to enhance the stability and in vivo anti-hypertensive effect of Ala-Leu-Gly-Pro-Gln-Phe-Tyr through encapsulation, thereby making it potentially valuable for various applications in pharmaceuticals and food industry. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
13. Evaluation of resazurin phenoxazine dye as a highly sensitive cell viability potency assay for natural killer cell‐derived extracellular vesicle‐based cancer biotherapeutics.
- Author
-
St‐Denis‐Bissonnette, Frederic, Qiu, Shirley, Cummings, Sarah E., Kirkby, Melanie, Haile, Yohannes, Wassmer, Sarah, Muradia, Gauri, Mehic, Jelica, Stalker, Andrew, Shrestha, Amit, Ardolino, Michele, Lee, Seung‐Hwan, Burger, Dylan, Wang, Lisheng, and Lavoie, Jessie R.
- Subjects
- *
CELL survival , *CANCER cell analysis , *RESAZURIN , *CYTOTOXINS , *EXTRACELLULAR vesicles , *LACTATE dehydrogenase - Abstract
Natural killer cell‐derived extracellular vesicles (NK‐EVs) are candidate biotherapeutics against various cancers. However, standardised potency assays are necessary for a reliable assessment of NK‐EVs' cytotoxicity. This study aims to thoroughly evaluate a highly sensitive resazurin phenoxazine‐based cell viability potency assay (measurement of the cellular redox metabolism) for quantifying the cytotoxicity of NK‐EVs against leukaemia K562 cells (suspension model) and breast cancer MDA‐MB‐231 cells (adherent model) in vitro. The assay was evaluated based on common analytical parameters setforth by regulatory guidelines, including specificity, selectivity,accuracy, precision, linearity, range and stability. Our results revealed that this resazurin‐based cell viability potency assay reliably and reproducibly measured a dose‐response of NK‐EVs' cytotoxic activity against both cancer models. The assay showed precision with 5% and 20% variation for intra‐run and inter‐run variability. The assay signal showed specificity and selectivity of NK‐EVs against cancer target cells, as evidenced by the diminished viability of cancer cells following a 5‐hour treatment with NK‐EVs, without any detectable interference or background. The linearity analysis of target cancer cells revealed strong linearity for densities of 5000 K562 and 1000 MDA‐MB‐231 cells per test with a consistent range. Importantly, NK‐EVs' dose‐response for cytotoxicity showed a strong correlation (|ρ| ∼ 0.8) with the levels of known cytotoxic factors associated with the NK‐EVs' corona (FasL, GNLY, GzmB, PFN and IFN‐γ), thereby validating the accuracy of the assay. The assay also distinguished cytotoxicity changes in degraded NK‐EVs, indicating the ability of the assay to detect the potential loss of sample integrity. Compared to other commonly reported bioassays (i.e., flow cytometry, cell counting, lactate dehydrogenase release assay, DNA‐binding reporter assay and confluence assay), our results support this highly sensitive resazurin‐based viability potency assay as a high‐throughput and quantitative method for assessing NK‐EVs' cytotoxicity against both suspension and adherent cancer models for evaluating NK‐EVs' biotherapeutics. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
14. Global outlook on affordability of biotherapeutic drugs.
- Author
-
Rathore, Anurag S., Gardner, Peter J., Chhabra, Hemlata, and Raman, Ruchir
- Subjects
- *
LOW-income countries , *MIDDLE-income countries , *HIGH-income countries , *PRICES , *WORLD health - Abstract
Although biotherapeutic drugs have the potential of transforming the management of many life‐threatening diseases, their affordability and accessibility remain an issue. This study offers an overview of the global affordability of biotherapeutic products. For this, prices for 10 representative biotherapeutic products were examined in 40 countries, including high‐income countries (HICs), upper middle‐income countries (UMICs), lower middle‐income countries (LMICs), and low‐income countries (LICs). The affordability of these biotherapeutics was calculated based on the World Health Organization/Health Action International (WHO/HAI) method. As expected, affordability was found to be better in HICs, followed by UMICs, LMICs, and finally, LICs. Furthermore, based on the trend of per capita income, we predict that in UMICs and LMICs, the affordability of high molecular weight biologics will worsen by 1.5× and 2× by 2030, respectively, and further by 4× and 6× by 2040. On the other hand, affordability will stay nearly the same for people living in HICs in the coming decades. Our analysis suggests that it is imperative that measures be taken to make this class of products more affordable and accessible. Governments can contribute by creating conducive policies. Global institutions like the WHO can play a significant role as well. Finally, manufacturers need to invest in and implement manufacturing innovations. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
15. Bioinspired and bioderived nanomedicine for inflammatory bowel disease.
- Author
-
Gazzi, Rafaela, Gelli, Rita, Aleandri, Simone, Carone, Marianna, and Luciani, Paola
- Abstract
Due to its chronic nature and complex pathophysiology, inflammatory bowel disease (IBD) poses significant challenges for treatment. The long‐term therapies for patients, often diagnosed between the ages of 20 and 40, call for innovative strategies to target inflammation, minimize systemic drug exposure, and improve patients' therapeutic outcomes. Among the plethora of strategies currently pursued, bioinspired and bioderived nano‐based formulations have garnered interest for their safety and versatility in the management of IBD. Bioinspired nanomedicine can host and deliver not only small drug molecules but also biotherapeutics, be made gastroresistant and mucoadhesive or mucopenetrating and, for these reasons, are largely investigated for oral administration, while surprisingly less for rectal delivery, recommended first‐line treatment approach for several IBD patients. The use of bioderived nanocarriers, mostly extracellular vesicles (EVs), endowed with unique homing abilities, is still in its infancy with respect to the arsenal of nanomedicine under investigation for IBD treatment. An emerging source of EVs suited for oral administration is ingesta, that is, plants or milk, thanks to their remarkable ability to resist the harsh environment of the upper gastrointestinal tract. Inspired by the unparalleled properties of natural biomaterials, sophisticated avenues for enhancing therapeutic efficacy and advancing precision medicine approaches in IBD care are taking shape, although bottlenecks arising either from the complexity of the nanomedicine designed or from the lack of a clear regulatory pathway still hinder a smooth and efficient translation to the clinics. This article is categorized under:Nanotechnology Approaches to Biology > Nanoscale Systems in Biology [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
16. Transdermal Administration of Nanobody Molecules using Hydrogel‐Forming Microarray Patch Technology: A Unique Delivery Approach.
- Author
-
Hutton, Aaron R. J., Kirkby, Melissa, Van Bogaert, Tom, Casteels, Peter, Nonne, Christelle, De Brabandere, Veronique, de Vyver, Ortwin Van, Vora, Lalit K., Tekko, Ismaiel A., McCarthy, Helen O., and Donnelly, Ryan F.
- Subjects
- *
MICROARRAY technology , *MULTIVALENT molecules , *HYPODERMIC needles , *SPRAGUE Dawley rats , *MOLECULES - Abstract
Nanobody molecules, derived from heavy‐chain only antibodies in camelids, represent the next generation of biotherapeutics. In addition to low immunogenicity, high stability, and potency, their single‐domain format facilitates the construction of multivalent molecules for therapeutic applications. Although predominantly administered using a hypodermic syringe and needle, alternative delivery methods are under investigation. That said, the transdermal route has yet to be explored. Therefore, microarray patch (MAP) technology, offering a potentially high dose, pain‐free transdermal system, is employed in this study. Trivalent Nanobody molecules, with and without half‐life extension (VHH and VHH[HLE]), are formulated into hydrogel‐forming MAPs, with pharmacokinetic parameters assessed in Sprague–Dawley rats. VHH MAPs exhibited a sustained release profile, with a serum concentration of 19 ± 9 ng mL−1 24 h post‐administration. In contrast, a subcutaneous (SC) injection showed faster clearance, with a serum concentration of 1.1 ± 0.4 ng mL−1 at 24 h. For VHH(HLE), both SC and MAP cohorts achieved a maximum serum concentration (Tmax) at 24 h. The MAP cohort displayed a notable increase in VHH(HLE) serum levels between 6–24 h, dropping after MAP removal. This study has exemplified MAPs potential for delivering advanced biologics, indicating the transdermal route's promise for pain‐free, patient‐friendly administration of Nanobody molecules. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
17. Liquid-phase separations coupled with ion mobility-mass spectrometry for next-generation biopharmaceutical analysis.
- Author
-
Makey, Devin M. and Ruotolo, Brandon T.
- Abstract
The pharmaceutical industry continues to expand its search for innovative biotherapeutics. The comprehensive characterization of such therapeutics requires many analytical techniques to fully evaluate critical quality attributes, making analysis a bottleneck in discovery and development timelines. While thorough characterization is crucial for ensuring the safety and efficacy of biotherapeutics, there is a need to further streamline analytical characterization and expedite the overall timeline from discovery to market. This review focuses on recent developments in liquid-phase separations coupled with ion mobility-mass spectrometry (IM-MS) for the development and characterization of biotherapeutics. We cover uses of IM-MS to improve the characterization of monoclonal antibodies, antibody-drug conjugates, host cell proteins, glycans, and nucleic acids. This discussion is based on an extensive literature search using Web of Science, Google Scholar, and SciFinder. IM-MS has the potential to enhance the depth and efficiency of biotherapeutic characterization by providing additional insights into conformational changes, post-translational modifications, and impurity profiles. The rapid timescale of IM-MS positions it well to enhance the information content of existing assays through its facile integration with standard liquid-phase separation techniques that are commonly used for biopharmaceutical analysis. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
18. Internalization of therapeutic antibodies into dendritic cells as a risk factor for immunogenicity
- Author
-
Michel Siegel, Anna-Lena Bolender, Axel Ducret, Johannes Fraidling, Katharina Hartman, Cary M. Looney, Olivier Rohr, Timothy P. Hickling, Hubert Kettenberger, Martin Lechmann, Céline Marban-Doran, and Thomas E. Kraft
- Subjects
immunogenicity ,biotherapeutics ,charge patches ,internalization ,dendritic cells ,Immunologic diseases. Allergy ,RC581-607 - Abstract
IntroductionImmunogenicity, the unwanted immune response triggered by therapeutic antibodies, poses significant challenges in biotherapeutic development. This response can lead to the production of anti-drug antibodies, potentially compromising the efficacy and safety of treatments. The internalization of therapeutic antibodies into dendritic cells (DCs) is a critical factor influencing immunogenicity. Using monoclonal antibodies, with differences in non-specific cellular uptake, as tools to explore the impact on the overall risk of immunogenicity, this study explores how internalization influences peptide presentation and subsequently T cell activation.Materials and methodsTo investigate the impact of antibody internalization on immunogenicity, untargeted toolantibodies with engineered positive or negative charge patches were utilized. Immature monocyte-derived DCs (moDCs), known for their physiologically relevant high endocytic activity, were employed for internalization assays, while mature moDCs were used for MHC-II associated peptide proteomics (MAPPs) assays. In addition to the lysosomal accumulation and peptide presentation, subsequent CD4+ T cell activation has been assessed. Consequently, a known CD4+ T cell epitope from ovalbumin was inserted into the tool antibodies to evaluate T cell activation on a single, shared epitope.ResultsAntibodies with positive charge patches exhibited higher rates of lysosomal accumulation and epitope presentation compared to those with negative charge patches or neutral surface charge. Furthermore, a direct correlation between internalization rate and presentation on MHC-II molecules could be established. To explore the link between internalization, peptide presentation and CD4+ T cell activation, tool antibodies containing the same OVA epitope were used. Previous observations were not altered by the insertion of the OVA epitope and ultimately, an enhanced CD4+ T cell response correlated with increased internalization in DCs and peptide presentation.DiscussionThese findings demonstrate that the biophysical properties of therapeutic antibodies, particularly surface charge, play a crucial role in their internalization into DCs. Antibodies internalized faster and processed by DCs, are also more prone to be presented on their surface leading to a higher risk of triggering an immune response. These insights underscore the importance of considering antibody surface charge and other properties that enhance cellular accumulation during the preclinical development of biotherapeutics to mitigate immunogenicity risks.
- Published
- 2024
- Full Text
- View/download PDF
19. Development and characterization of dendritic cell internalization and activation assays contributing to the immunogenicity risk evaluation of biotherapeutics
- Author
-
Michel Siegel, Aman Padamsey, Anna-Lena Bolender, Patrick Hargreaves, Johannes Fraidling, Axel Ducret, Katharina Hartman, Cary M. Looney, Cristina Bertinetti-Lapatki, Olivier Rohr, Timothy P. Hickling, Thomas E. Kraft, and Céline Marban-Doran
- Subjects
immunogenicity ,immunomodulation ,biotherapeutics ,dendritic cells ,assay development ,Immunologic diseases. Allergy ,RC581-607 - Abstract
IntroductionImmunogenicity refers to the ability of a substance, such as a therapeutic drug, to elicit an immune response. While beneficial in vaccine development, undesirable immunogenicity can compromise the safety and efficacy of therapeutic proteins by inducing anti-drug antibodies (ADAs). These ADAs can reduce drug bioavailability and alter pharmacokinetics, necessitating comprehensive immunogenicity risk assessments starting at early stages of drug development. Given the complexity of immunogenicity, an integrated approach is essential, as no single assay can universally recapitulate the immune response leading to the formation of anti-drug antibodies.MethodsTo better understand the Dendritic Cell (DC) contribution to immunogenicity, we developed two flow cytometry-based assays: the DC internalization assay and the DC activation assay. Monocyte-derived dendritic cells (moDCs) were generated from peripheral blood mononuclear cells (PBMCs) and differentiated over a five-day period. The internalization assay measured the accumulation rate of therapeutic antibodies within moDCs, while the activation assay assessed the expression of DC activation markers such as CD40, CD80, CD86, CD83, and DC-SIGN (CD209). To characterize these two assays further, we used a set of marketed therapeutic antibodies.ResultsThe study highlights that moDCs differentiated for 5 days from freshly isolated monocytes were more prone to respond to external stimuli. The internalization assay has been shown to be highly sensitive to the molecule tested, allowing the use of only 4 donors to detect small but significant differences. We also demonstrated that therapeutic antibodies were efficiently taken up by moDCs, with a strong correlation with their peptide presentation on MHC-II. On the other hand, by monitoring DC activation through a limited set of activation markers including CD40, CD83, and DC-SIGN, the DC activation assay has the potential to compare a series of compounds. These two assays provide a more comprehensive understanding of DC function in the context of immunogenicity, highlighting the importance of both internalization and activation processes in ADA development.DiscussionThe DC internalization and activation assays described here address key gaps in existing immunogenicity assessment methods by providing specific and reliable measures of DC function. The assays enhance our ability to pre-clinically evaluate the immunogenic potential of biotherapeutics, thereby improving their safety and efficacy. Future work should focus on further validating these assays and integrating them into a holistic immunogenicity risk assessment framework.
- Published
- 2024
- Full Text
- View/download PDF
20. Metabolic Materials from Cyanobacteria and Microalgae: Biotechnology, Biochemistry and Biotherapeutics
- Author
-
Paduvetnaya, Lokitha, Venkatesh, Kamath H., Madhyastha, Harishkumar, Agarwal, Avinash Kumar, Series Editor, Bala, Kiran, editor, Ghosh, Tonmoy, editor, Kumar, Vivek, editor, and Sangwan, Pritam, editor
- Published
- 2024
- Full Text
- View/download PDF
21. Microneedle and Polymeric Films: Delivery of Proteins, Peptides and Nucleic Acids
- Author
-
Wu, Yu, Hutton, Aaron R. J., Pandya, Anjali Kiran, Patravale, Vandana B., Donnelly, Ryan F., Michel, Martin C., Editor-in-Chief, Barrett, James E., Editorial Board Member, Centurión, David, Editorial Board Member, Flockerzi, Veit, Editorial Board Member, Meier, Kathryn Elaine, Editorial Board Member, Page, Clive P., Editorial Board Member, Seifert, Roland, Editorial Board Member, Wang, KeWei, Editorial Board Member, Schäfer-Korting, Monika, editor, and Schubert, Ulrich S., editor
- Published
- 2024
- Full Text
- View/download PDF
22. A novel front in sustainable microbial management: computational analysis of curcumin and mangiferin's synergistic action against Bacillus anthracis.
- Author
-
Perveen, Kahkashan, Bukhari, Najat A., Alshaikh, Najla A., Kondaveeti, Suresh Babu, Alsulami, Jamilah A., Debnath, Sandip, and Kumarasamy, Vinoth
- Subjects
BACILLUS anthracis ,MANGIFERIN ,CURCUMIN ,MOLECULAR dynamics ,BINDING energy - Abstract
Background: Microorganisms are crucial in our ecosystem, offering diverse functions and adaptability. The UNGA Science Summit has underscored the importance of understanding microbes in alignment with the UN Sustainable Development Goals. Bacillus anthracis poses significant challenges among various microorganisms due to its harmful effects on both soil and public health. Our study employed computational techniques to investigate the inhibitory effects of curcumin and mangiferin on Bacillus anthracis, with the aim of presenting a novel bio-based approach to microbial management. Methods: Employing high-throughput screening, we identified potential binding sites on B. anthracis. Molecular docking revealed that curcumin and mangiferin, when synergistically combined, exhibited strong binding affinities at different sites on the bacterium. Our findings demonstrated a significant drop in binding free energy, indicating a stronger interaction when these compounds were used together. Findings: Results of Molecular docking indicated binding energies of -8.45 kcal/mol for mangiferin, -7.68 kcal/mol for curcumin, and a notably higher binding energy of -19.47 kcal/mol for the combination of mangiferin and curcumin with CapD protein. Molecular dynamics simulations further validated these interactions, demonstrating increased stability and structural changes in the bacterium. Conclusion: This study highlights the effectiveness of natural compounds like curcumin and mangiferin in microbial management, especially against challenging pathogens like B. anthracis. It emphasizes the potential of sustainable, nature-based solutions and calls for further empirical research to expand upon these findings. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
23. Prophylactic treatment with PEGylated bovine IFNλ3 effectively bridges the gap in vaccine-induced immunity against FMD in cattle.
- Author
-
Attreed, Sarah E., Silva, Christina, Rodriguez-Calzada, Monica, Mogulothu, Aishwarya, Abbott, Sophia, Azzinaro, Paul, Canning, Peter, Skidmore, Lillian, Nelson, Jay, Knudsen, Nick, Medina, Gisselle N., de los Santos, Teresa, and Segundo, Fayna Díaz-San
- Subjects
CATTLE ,COMBINED vaccines ,BOS ,ANIMAL diseases ,VIRUS diseases ,FOOT & mouth disease - Abstract
Foot-and-mouth disease (FMD) is a vesicular disease of cloven-hoofed animals with devastating economic implications. The current FMD vaccine, routinely used in enzootic countries, requires at least 7 days to induce protection. However, FMD vaccination is typically not recommended for use in non-enzootic areas, underscoring the need to develop new fast-acting therapies for FMD control during outbreaks. Interferons (IFNs) are among the immune system's first line of defense against viral infections. Bovine type III IFN delivered by a replication defective adenovirus (Ad) vector has effectively blocked FMD in cattle. However, the limited duration of protection--usually only 1-3 days post-treatment (dpt)--diminishes its utility as a field therapeutic. Here, we test whether polyethylene glycosylation (PEGylation) of recombinant bovine IFNλ3 (PEGboIFNλ3) can extend the duration of IFN-induced prevention of FMDV infection in both vaccinated and unvaccinated cattle. We treated groups of heifers with PEGboIFNλ3 alone or in combination with an adenovirus-based FMD O1Manisa vaccine (Adt-O1M) at either 3 or 5 days prior to challenge with homologous wild type FMDV. We found that pre-treatment with PEGboIFNλ3 was highly effective at preventing clinical FMD when administered at either time point, with or without co-administration of Adt-O1M vaccine. PEGboIFNλ3 protein was detectable systemically for >10 days and antiviral activity for 4 days following administration. Furthermore, in combination with Adt-O1M vaccine, we observed a strong induction of FMDV-specific IFNγ+ T cell response, demonstrating its adjuvanticity when co-administered with a vaccine. Our results demonstrate the promise of this modified IFN as a pre-exposure prophylactic therapy for use in emergency outbreak scenarios. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
24. The effect of the composition of polysorbate 80 grades on their physicochemical properties.
- Author
-
Kollamaram, Gayathri and Williams, Gareth R.
- Subjects
- *
POLYSORBATE 80 , *FATTY acids , *EXCIPIENTS - Abstract
Polysorbate 80 is one of the most commonly used surfactants in the formulation of biotherapeutics, particularly those administered intravenously. It comprises a mixture of fatty acids but is not a precisely defined chemical entity. Hence, there are a range of different grades available in the market, all meeting compendial specifications. Polysorbate 80 is known to undergo auto degradation producing protein-damaging by-products, and to contain residual impurities that can have an impact on the stability and integrity of the active ingredients in the formulation. Given the variety of chemical compositions that polysorbate 80 can comprise, the degradation pathway and extent could vary depending on the grade used in the formulation. This study compared the physical and chemical properties of four commercially available polysorbate 80 grades with different degrees of purity and oleic acid content and investigated their degradation profiles. This study did not find any significant differences between the properties or degradation profiles of the four grades investigated. Further studies are underway to understand the formation of other reactive impurities and their impact on the model protein formulations. [ABSTRACT FROM AUTHOR]
- Published
- 2024
25. The NMR signature of maltose-based glycation in full-length proteins.
- Author
-
Defant, Pauline, Regl, Christof, Huber, Christian G., and Schubert, Mario
- Subjects
POST-translational modification ,PROTEINS ,NUCLEAR magnetic resonance spectroscopy ,MALTOSE ,DISACCHARIDES - Abstract
Reducing sugars can spontaneously react with free amines in protein side chains leading to posttranslational modifications (PTMs) called glycation. In contrast to glycosylation, glycation is a non-enzymatic modification with consequences on the overall charge, solubility, aggregation susceptibility and functionality of a protein. Glycation is a critical quality attribute of therapeutic monoclonal antibodies. In addition to glucose, also disaccharides like maltose can form glycation products. We present here a detailed NMR analysis of the Amadori product formed between proteins and maltose. For better comparison, data collection was done under denaturing conditions using 7 M urea-d
4 in D2 O. The here presented correlation patterns serve as a signature and can be used to identify maltose-based glycation in any protein that can be denatured. In addition to the model protein BSA, which can be readily glycated, we present data of the biotherapeutic abatacept containing maltose in its formulation buffer. With this contribution, we demonstrate that NMR spectroscopy is an independent method for detecting maltose-based glycation, that is suited for cross-validation with other methods. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
26. The Microbiome and Pediatric Transplantation.
- Author
-
Elgarten, Caitlin W, Margolis, Elisa B, and Kelly, Matthew S
- Subjects
- *
INFECTION risk factors , *RISK assessment , *IMMUNOLOGICAL tolerance , *GRAFT versus host disease , *HEMATOPOIETIC stem cell transplantation , *TRANSPLANTATION of organs, tissues, etc. , *GUT microbiome , *GRAFT rejection , *CHILDREN - Abstract
The microbial communities that inhabit our bodies have been increasingly linked to host physiology and pathophysiology. This microbiome, through its role in colonization resistance, influences the risk of infections after transplantation, including those caused by multidrug-resistant organisms. In addition, through both direct interactions with the host immune system and via the production of metabolites that impact local and systemic immunity, the microbiome plays an important role in the establishment of immune tolerance after transplantation, and conversely, in the development of graft-versus-host disease and graft rejection. This review offers a comprehensive overview of the evidence for the role of the microbiome in hematopoietic cell and solid organ transplant complications, drivers of microbiome shift during transplantation, and the potential of microbiome-based therapies to improve pediatric transplantation outcomes. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
27. Evaluation of resazurin phenoxazine dye as a highly sensitive cell viability potency assay for natural killer cell‐derived extracellular vesicle‐based cancer biotherapeutics
- Author
-
Frederic St‐Denis‐Bissonnette, Shirley Qiu, Sarah E. Cummings, Melanie Kirkby, Yohannes Haile, Sarah Wassmer, Gauri Muradia, Jelica Mehic, Andrew Stalker, Amit Shrestha, Michele Ardolino, Seung‐Hwan Lee, Dylan Burger, Lisheng Wang, and Jessie R. Lavoie
- Subjects
biotherapeutics ,cancer ,exosome ,extracellular vesicles (EVs) ,natural killer (NK) cells ,potency assay ,Cytology ,QH573-671 - Abstract
Abstract Natural killer cell‐derived extracellular vesicles (NK‐EVs) are candidate biotherapeutics against various cancers. However, standardised potency assays are necessary for a reliable assessment of NK‐EVs' cytotoxicity. This study aims to thoroughly evaluate a highly sensitive resazurin phenoxazine‐based cell viability potency assay (measurement of the cellular redox metabolism) for quantifying the cytotoxicity of NK‐EVs against leukaemia K562 cells (suspension model) and breast cancer MDA‐MB‐231 cells (adherent model) in vitro. The assay was evaluated based on common analytical parameters setforth by regulatory guidelines, including specificity, selectivity,accuracy, precision, linearity, range and stability. Our results revealed that this resazurin‐based cell viability potency assay reliably and reproducibly measured a dose‐response of NK‐EVs’ cytotoxic activity against both cancer models. The assay showed precision with 5% and 20% variation for intra‐run and inter‐run variability. The assay signal showed specificity and selectivity of NK‐EVs against cancer target cells, as evidenced by the diminished viability of cancer cells following a 5‐hour treatment with NK‐EVs, without any detectable interference or background. The linearity analysis of target cancer cells revealed strong linearity for densities of 5000 K562 and 1000 MDA‐MB‐231 cells per test with a consistent range. Importantly, NK‐EVs’ dose‐response for cytotoxicity showed a strong correlation (|ρ| ∼ 0.8) with the levels of known cytotoxic factors associated with the NK‐EVs’ corona (FasL, GNLY, GzmB, PFN and IFN‐γ), thereby validating the accuracy of the assay. The assay also distinguished cytotoxicity changes in degraded NK‐EVs, indicating the ability of the assay to detect the potential loss of sample integrity. Compared to other commonly reported bioassays (i.e., flow cytometry, cell counting, lactate dehydrogenase release assay, DNA‐binding reporter assay and confluence assay), our results support this highly sensitive resazurin‐based viability potency assay as a high‐throughput and quantitative method for assessing NK‐EVs’ cytotoxicity against both suspension and adherent cancer models for evaluating NK‐EVs’ biotherapeutics.
- Published
- 2024
- Full Text
- View/download PDF
28. Transdermal Administration of Nanobody Molecules using Hydrogel‐Forming Microarray Patch Technology: A Unique Delivery Approach
- Author
-
Aaron R. J. Hutton, Melissa Kirkby, Tom Van Bogaert, Peter Casteels, Christelle Nonne, Veronique De Brabandere, Ortwin Van deVyver, Lalit K. Vora, Ismaiel A. Tekko, Helen O. McCarthy, and Ryan F. Donnelly
- Subjects
biotherapeutics ,hydrogel‐forming ,microarray patch ,transdermal ,Materials of engineering and construction. Mechanics of materials ,TA401-492 ,Engineering (General). Civil engineering (General) ,TA1-2040 - Abstract
Abstract Nanobody molecules, derived from heavy‐chain only antibodies in camelids, represent the next generation of biotherapeutics. In addition to low immunogenicity, high stability, and potency, their single‐domain format facilitates the construction of multivalent molecules for therapeutic applications. Although predominantly administered using a hypodermic syringe and needle, alternative delivery methods are under investigation. That said, the transdermal route has yet to be explored. Therefore, microarray patch (MAP) technology, offering a potentially high dose, pain‐free transdermal system, is employed in this study. Trivalent Nanobody molecules, with and without half‐life extension (VHH and VHH[HLE]), are formulated into hydrogel‐forming MAPs, with pharmacokinetic parameters assessed in Sprague–Dawley rats. VHH MAPs exhibited a sustained release profile, with a serum concentration of 19 ± 9 ng mL−1 24 h post‐administration. In contrast, a subcutaneous (SC) injection showed faster clearance, with a serum concentration of 1.1 ± 0.4 ng mL−1 at 24 h. For VHH(HLE), both SC and MAP cohorts achieved a maximum serum concentration (Tmax) at 24 h. The MAP cohort displayed a notable increase in VHH(HLE) serum levels between 6–24 h, dropping after MAP removal. This study has exemplified MAPs potential for delivering advanced biologics, indicating the transdermal route's promise for pain‐free, patient‐friendly administration of Nanobody molecules.
- Published
- 2024
- Full Text
- View/download PDF
29. EMID2 is a novel biotherapeutic for aggressive cancers identified by in vivo screening
- Author
-
Ambra Cappelletto, Edoardo Alfì, Nina Volf, Thi Van Anh Vu, Francesca Bortolotti, Giulio Ciucci, Simone Vodret, Marco Fantuz, Martina Perin, Andrea Colliva, Giacomo Rozzi, Matilde Rossi, Giulia Ruozi, Lorena Zentilin, Roman Vuerich, Daniele Borin, Romano Lapasin, Silvano Piazza, Mattia Chiesa, Daniela Lorizio, Luca Triboli, Sandeep Kumar, Gaia Morello, Claudio Tripodo, Maurizio Pinamonti, Giulia Maria Piperno, Federica Benvenuti, Alessandra Rustighi, Hanjoong Jo, Stefano Piccolo, Giannino Del Sal, Alessandro Carrer, Mauro Giacca, and Serena Zacchigna
- Subjects
In vivo screening ,Cancer ,Cell invasiveness ,AAV vectors ,Gene therapy ,Biotherapeutics ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Abstract Background New drugs to tackle the next pathway or mutation fueling cancer are constantly proposed, but 97% of them are doomed to fail in clinical trials, largely because they are identified by cellular or in silico screens that cannot predict their in vivo effect. Methods We screened an Adeno-Associated Vector secretome library (> 1000 clones) directly in vivo in a mouse model of cancer and validated the therapeutic effect of the first hit, EMID2, in both orthotopic and genetic models of lung and pancreatic cancer. Results EMID2 overexpression inhibited both tumor growth and metastatic dissemination, consistent with prolonged survival of patients with high levels of EMID2 expression in the most aggressive human cancers. Mechanistically, EMID2 inhibited TGFβ maturation and activation of cancer-associated fibroblasts, resulting in more elastic ECM and reduced levels of YAP in the nuclei of cancer cells. Conclusion This is the first in vivo screening, precisely designed to identify proteins able to interfere with cancer cell invasiveness. EMID2 was selected as the most potent protein, in line with the emerging relevance of the tumor extracellular matrix in controlling cancer cell invasiveness and dissemination, which kills most of cancer patients.
- Published
- 2024
- Full Text
- View/download PDF
30. Pulmonary Inhalation of Biotherapeutics: A Systematic Approach to Understanding the Effects of Atomisation Gas Flow Rate on Particle Physiochemical Properties and Retained Bioactivity
- Author
-
Laura Foley, Ahmad Ziaee, Gavin Walker, and Emmet O’Reilly
- Subjects
biotherapeutics ,spray-drying ,atomisation gas flow rate ,enzymatic activity ,particle engineering ,pulmonary delivery ,Pharmacy and materia medica ,RS1-441 - Abstract
The identification of spray-drying processing parameters capable of producing particles suitable for pulmonary inhalation with retained bioactivity underpins the development of inhalable biotherapeutics. Effective delivery of biopharmaceuticals via pulmonary delivery routes such as dry powder inhalation (DPI) requires developing techniques that engineer particles to well-defined target profiles while simultaneously minimising protein denaturation. This study examines the simultaneous effects of atomisation gas flow rate on particle properties and retained bioactivity for the model biopharmaceutical lysozyme. The results show that optimising the interplay between atomisation gas flow rate and excipient concentration enables the production of free-flowing powder with retained bioactivity approaching 100%, moisture content below 4%, and D50 < 4 µm, at yields exceeding 50%. The developed methodologies inform the future design of protein-specific spray-drying parameters for inhalable biotherapeutics.
- Published
- 2024
- Full Text
- View/download PDF
31. Exploring the Multifaceted Therapeutic Potential of Probiotics: A Review of Current Insights and Applications
- Author
-
Chakravarty, Kashyapi, Gaur, Smriti, Kumar, Rohit, Jha, Niraj Kumar, and Gupta, Piyush Kumar
- Published
- 2024
- Full Text
- View/download PDF
32. Unveiling Curvularia tuberculata-induced leaf anomalies in Rhododendron ferrugineum: implications in cultural-ecological conservation and harnessing microbial intervention in socio-economic advancement.
- Author
-
Dhar, Juhita, Hazra, Aishee, Patra, Riddhisha, Kumar, Varun, Subramaniyan, Vetriselvan, Kumarasamy, Vinoth, Mitra, Arup Kumar, Sayed, Amany A., Aleya, Lotfi, El-Demerdash, Fatma M., Almutairi, Mikhlid H., Akash, Shopnil, Abdel-Daim, Mohamed M., Kant, Achal, and Dhara, Bikram
- Subjects
RHODODENDRONS ,CURVULARIA ,PATHOGENIC fungi ,MOUNTAIN ecology ,CULTURAL property ,DISEASE management - Abstract
Introduction: The research focuses on Rhododendron ferrugineum L., Nepal's national flower and Uttarakhand's state tree, thriving in high-altitude mountain ecosystems. Methodology and Result: A study conducted in Himachal Pradesh (Latitude: N 31° 6' 2.0088", Longitude: E 77° 10' 29.9136") identified leaf anomalies resembling rust-like manifestations in R. ferrugineum. These anomalies were traced back to the pathogenic fungus Curvularia tuberculata, marking the first documented case of its impact on R. ferrugineum in India. Discussion: This discovery emphasizes the need for vigilant monitoring, disease management research, and conservation efforts to protect the cultural and ecological significance of this iconic shrub. Beyond its immediate findings, the study introduces a novel dimension to Indian flora by associating C. tuberculata with R. ferrugineum, historically linked to monocotyledonous crops. The research methodology combines traditional microscopic examination with advanced genomic sequencing and phylogenetic analysis, enhancing pathogen identification accuracy. Future prospect: In a broader context, this research aligns with the United Nations Sustainable Development Goals (SDGs) by highlighting the importance of environmental preservation, conservation, and sustainable management. It underscores the intricate interplay between biodiversity, cultural heritage, and the need for holistic solutions. Overall, this study calls for proactive measures to protect R. ferrugineum's cultural and ecological heritage and emphasizes the significance of interdisciplinary approaches in addressing emerging ecological threats. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
33. EMID2 is a novel biotherapeutic for aggressive cancers identified by in vivo screening.
- Author
-
Cappelletto, Ambra, Alfì, Edoardo, Volf, Nina, Vu, Thi Van Anh, Bortolotti, Francesca, Ciucci, Giulio, Vodret, Simone, Fantuz, Marco, Perin, Martina, Colliva, Andrea, Rozzi, Giacomo, Rossi, Matilde, Ruozi, Giulia, Zentilin, Lorena, Vuerich, Roman, Borin, Daniele, Lapasin, Romano, Piazza, Silvano, Chiesa, Mattia, and Lorizio, Daniela
- Subjects
- *
MEDICAL screening , *GENETIC models , *CANCER invasiveness , *EXTRACELLULAR matrix , *CELL nuclei , *BIOSURVEILLANCE - Abstract
Background: New drugs to tackle the next pathway or mutation fueling cancer are constantly proposed, but 97% of them are doomed to fail in clinical trials, largely because they are identified by cellular or in silico screens that cannot predict their in vivo effect. Methods: We screened an Adeno-Associated Vector secretome library (> 1000 clones) directly in vivo in a mouse model of cancer and validated the therapeutic effect of the first hit, EMID2, in both orthotopic and genetic models of lung and pancreatic cancer. Results: EMID2 overexpression inhibited both tumor growth and metastatic dissemination, consistent with prolonged survival of patients with high levels of EMID2 expression in the most aggressive human cancers. Mechanistically, EMID2 inhibited TGFβ maturation and activation of cancer-associated fibroblasts, resulting in more elastic ECM and reduced levels of YAP in the nuclei of cancer cells. Conclusion: This is the first in vivo screening, precisely designed to identify proteins able to interfere with cancer cell invasiveness. EMID2 was selected as the most potent protein, in line with the emerging relevance of the tumor extracellular matrix in controlling cancer cell invasiveness and dissemination, which kills most of cancer patients. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
34. Enhancing Oral Delivery of Biologics: A Non-Competitive and Cross-Reactive Anti-Leptin Receptor Nanofitin Demonstrates a Gut-Crossing Capacity in an Ex Vivo Porcine Intestinal Model.
- Author
-
Masloh, Solene, Chevrel, Anne, Culot, Maxime, Perrocheau, Anaëlle, Kalia, Yogeshvar N., Frehel, Samuel, Gaussin, Rémi, Gosselet, Fabien, Huet, Simon, Zeisser Labouebe, Magali, and Scapozza, Leonardo
- Subjects
- *
LEPTIN receptors , *INTESTINAL barrier function , *INTESTINES , *INTESTINAL absorption , *BIOLOGICALS , *LEPTIN - Abstract
Biotherapeutics exhibit high efficacy in targeted therapy, but their oral delivery is impeded by the harsh conditions of the gastrointestinal (GI) tract and limited intestinal absorption. This article presents a strategy to overcome the challenges of poor intestinal permeability by using a protein shuttle that specifically binds to an intestinal target, the leptin receptor (LepR), and exploiting its capacity to perform a receptor-mediated transport. Our proof-of-concept study focuses on the characterization and transport of robust affinity proteins, known as Nanofitins, across an ex vivo porcine intestinal model. We describe the potential to deliver biologically active molecules across the mucosa by fusing them with the Nanofitin 1-F08 targeting the LepR. This particular Nanofitin was selected for its absence of competition with leptin, its cross-reactivity with LepR from human, mouse, and pig hosts, and its shuttle capability associated with its ability to induce a receptor-mediated transport. This study paves the way for future in vivo demonstration of a safe and efficient oral-to-systemic delivery of targeted therapies. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
35. Human Dose and Pharmacokinetic Predictions for Biologics at Boehringer Ingelheim: A Retrospective Analysis.
- Author
-
Grempler, Rolf, Ahlberg, Jennifer, Germovsek, Eva, Gupta, Priyanka, Li, Hua, Pilvankar, Minu, Sharma, Ashish, Stopfer, Peter, and Hansel, Steven
- Abstract
Introduction: Accurate predictions of pharmacokinetics and efficacious doses for biologics in humans are critical for selecting appropriate first-in-human starting doses and dose ranges and for estimating clinical material needs and cost of goods. This also impacts clinical feasibility, particularly for subcutaneously administered biologics. Methods: We performed a comprehensive comparison between predicted and observed clearances and doses in humans for a set of 22 biologic drugs developed at Boehringer Ingelheim (BI) over the last 2 decades. The analysis included biologics across three therapeutic areas comprising a wide variety of modalities: mono- and bispecific monoclonal antibodies (mAbs) and nanobodies and a Fab fragment. Results: Our analysis showed that observed clearances in humans were within twofold of predicted clearances for 17 out of 20 biologics (85%). Six biologics had uncharacteristically high observed human clearances (range 32–280 mL/h) for their respective molecular classes, impacting their clinical developability. For three molecules, molecular characteristics contributed to the high clearance. Clinically selected doses were within twofold of predicted for 58% of projects. With 42% and 25% of projects selecting clinical doses higher than two- or threefold the predicted value, respectively, the importance of better understanding not only the pharmacokinetic (PK) but also the predictivity of pharmacodynamic models is highlighted. Conclusions: We provide a clinical pharmacology perspective on the commonly accepted twofold range of human clearance predictions as well as the implications of higher than predicted targeted efficacious plasma concentration on clinical development. Finally, an analysis of key success factors for biologics at BI was conducted, which may be relevant for the entire pharmaceutical industry. This is one of the largest retrospective analyses for biologics and provides further evidence that successful predictions of human PK and efficacious dose will be further facilitated by gathering key translational data early in research. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
36. Designer probiotics: Opening the new horizon in diagnosis and prevention of human diseases.
- Author
-
Debnath, Nabendu, Yadav, Pooja, Mehta, Praveen K., Gupta, Priyamvada, Kumar, Deepak, Kumar, Ashwani, Gautam, Vibhav, and Yadav, Ashok K.
- Abstract
Probiotic microorganisms have been used for therapeutic purposes for over a century, and recent advances in biotechnology and genetic engineering have opened up new possibilities for developing therapeutic approaches using indigenous probiotic microorganisms. Diseases are often related to metabolic and immunological factors, which play a critical role in their onset. With the help of advanced genetic tools, probiotics can be modified to produce or secrete important therapeutic peptides directly into mucosal sites, increasing their effectiveness. One potential approach to enhancing human health is through the use of designer probiotics, which possess immunogenic characteristics. These genetically engineered probiotics hold promise in providing novel therapeutic options. In addition to their immunogenic properties, designer probiotics can also be equipped with sensors and genetic circuits, enabling them to detect a range of diseases with remarkable precision. Such capabilities may significantly advance disease diagnosis and management. Furthermore, designer probiotics have the potential to be used in diagnostic applications, offering a less invasive and more cost‐effective alternative to conventional diagnostic techniques. This review offers an overview of the different functional aspects of the designer probiotics and their effectiveness on different diseases and also, we have emphasized their limitations and future implications. A comprehensive understanding of these functional attributes may pave the way for new avenues of prevention and the development of effective therapies for a range of diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
37. Alternatives to Monkey Reproductive Toxicology Testing for Biotherapeutics.
- Author
-
Hoberman, Alan M., Maki, Kazushige, Mikashima, Fumito, Naota, Misaki, Wange, Ronald L., Lansita, Janice A., and Weis, Shawna L.
- Subjects
- *
REPRODUCTIVE toxicology , *TOXICITY testing , *DEVELOPMENTAL toxicology , *BIOPHARMACEUTICS , *VETERINARY drugs , *MONKEYS , *KRA - Abstract
Embryofetal toxicity studies are conducted to support inclusion of women of childbearing potential in clinical trials and to support labeling for the marketed pharmaceutical product. For biopharmaceuticals, which frequently lack activity in the rodent or rabbit, the nonhuman primate is the standard model to evaluate embryofetal toxicity. These studies have become increasingly challenging to conduct due to the small number of facilities capable of performing them and a shortage of sexually mature monkeys. The low number of animals per group and the high rate of spontaneous abortion in cynomolgus monkeys further complicate interpretation of the data. Recent FDA guidance has proposed a weight of evidence (WoE) approach to support product labeling for reproductive toxicity of products intended to be used for the treatment of cancer (Oncology Pharmaceuticals: Reproductive Toxicity Testing and Labeling Recommendations), an approach that has also supported the approval of biotherapeutics for non-cancer indications. Considerations to determine the appropriateness and content of a WoE approach to support product labeling for embryofetal risk include known class effects in humans; findings from genetically modified animals with or without drug administration; information from surrogate compounds; literature-based assessments about the developmental role of the pharmaceutical target; and the anticipated exposure during embryofetal development. This paper summarizes the content of a session presented at the 42nd annual meeting at the American College of Toxicology, which explored the conditions under which alternative approaches may be appropriate to support product labeling for reproductive risk, and how sponsors can best justify the use of this approach. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
38. Emerging themes and factors influencing the prices of biotherapeutics.
- Author
-
Gandhi, Sonia, Kashiramka, Smita, and Rathore, Anurag S.
- Subjects
- *
PRICES , *DELIVERY of goods , *PRICE regulation , *PROFIT margins , *GOVERNMENT policy - Abstract
The high prices of biotherapeutics remain an area of concern for various national and international agencies working toward developing strategies for monitoring, evaluating, and managing the prices of medicines to promote equitable access. This paper deploys a unique study design to identify emerging themes and determine the elements influencing the price of biotherapeutics specifically biosimilars. This review is based on a bibliometric and qualitative synthesis approach to generate a new line of thinking for future studies. The findings enable knowledge assimilation for policy decisions. The results indicate that while the factors like market share, market competition, and pricing policies are extensively explored by researchers, the factors such as innovation capability, product development costs, and local factors like regulatory ecosystem can be further examined as potential influencers of prices. Further, pricing frameworks and regulations designed for generic drugs cannot be replicated to estimate the prices of biotherapeutics. As a result, policymakers must take the necessary actions to construct innovative pricing frameworks for this class of products that are in conformity with national policies, regulations, market dynamics, and economics. Moreover, studies on incentives and profit margins associated with the last‐mile delivery and accessibility of high‐cost biotherapeutics are essential for their efficient adoption. Key points: The high expense associated with the use of biotherapeutics, and the biosimilars that are used to treat cancer in particular, is a concern for many healthcare organizations.To strategize the relevant healthcare interventions, it is important to understand and synthesize the outcome of prior research on the price determinants of biotherapeutics, so as to facilitate future policy development.The national and international regulatory and health bodies can use the pricing factors and recommendations made via this research to create effective mechanisms for assessing and regulating the costs of expensive medicines like biosimilars.This article demonstrates how a combination of quantitative and qualitative methods can be used to review the corpus of research, resulting in the identification of important themes and variables affecting the cost of biotherapeutics.To comprehend the significant primary and secondary factors influencing the cost of biosimilars, the authors proposed a conceptual framework as a precursor for conducting evidence‐based investigations. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
39. Human secretory and excretory fluids, molecular constituents, and their biotherapeutic perspective against fungal pathogen Candida albicans.
- Author
-
Jaiswal, Neha and Kumar, Awanish
- Abstract
Candida albicans is a common fungal pathogen that can cause a range of infections, from superficial mucosal infections to invasive systemic infections, particularly in individuals with immunocompromised systems. The antimicrobial potential of human secretory and excretory fluids is known; however, relatively less attention has been paid to their antifungal potential. Therefore, it is important to understand the molecular constituents and antifungal properties of human secretory and excretory fluids against human fungal pathogen C. albicans to develop new strategies for combating the disease Candidiasis and enhance our understanding of the complex interplay between the host and pathogen. In this review, we discuss the potential antifungal molecules found in human saliva, vaginal fluid, cerumen, and urine. We also discussed the potential mechanisms of action of various antimicrobial peptides found in sweat, such as dermcidin and cathelicidin, and their potential as therapeutic agents against fungal infections. We have reviewed the information on anti-candidal molecules present in cerebrospinal fluid, cerumen, lacrimal fluid, sIgA, intestinal mucous, milk, saliva, seminal fluid, sweat, urine, etc. that would be very useful in a clinical context and futuristic biotherapeutic antifungal molecules. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
40. Development of a semi-automated MHC-associated peptide proteomics (MAPPs) method using streptavidin bead-based immunoaffinity capture and nano LC-MS/MS to support immunogenicity risk assessment in drug development.
- Author
-
Lee, M. Violet, Saad, Ola M., Wong, Sylvia, LaMar, Jason, Kamen, Lynn, Ordonia, Ben, Melendez, Rachel, Hassanzadeh, Azadeh, Shan Chung, and Kaur, Surinder
- Subjects
STREPTAVIDIN ,PEPTIDES ,IMMUNE response ,PROTEOMICS ,DRUG development ,MAJOR histocompatibility complex - Abstract
Major histocompatibility complex (MHC)-Associated Peptide Proteomics (MAPPs) is an ex vivo method used to assess the immunogenicity risk of biotherapeutics. MAPPs can identify potential T-cell epitopes within the biotherapeutic molecule. Using adalimumab treated human monocyte derived dendritic cells (DCs) and a pan anti-HLA-DR antibody (Ab), we systematically automated and optimized biotin/streptavidin (SA)-capture antibody coupling, lysate incubation with capture antibody, as well as the washing and elution steps of a MAPPs method using functionalized magnetic beads and a KingFisher Magnetic Particle processor. Automation of these steps, combined with capturing using biotinylated-Ab/SA magnetic beads rather than covalently bound antibody, improved reproducibility as measured by minimal inter-and intra-day variability, as well as minimal analyst-to-analyst variability. The semiautomated MAPPs workflow improved sensitivity, allowing for a lower number of cells per analysis. The method was assessed using five different biotherapeutics with varying immunogenicity rates ranging from 0.1 to 48% ADA incidence in the clinic. Biotherapeutics with ≥10%immunogenicity incidence consistently presented more peptides (1.8-28 fold) and clusters (10-21 fold) compared to those with <10% immunogenicity incidence. Our semi-automated MAPPs method provided two main advantages over a manual workflow- the robustness and reproducibility affords confidence in the epitopes identified from as few as 5 to 10 donors and the method workflow can be readily adapted to incorporate different capture Abs in addition to anti-HLA-DR. The incorporation of semi-automated MAPPs with biotinylated-Ab/SA bead-based capture in immunogenicity screening strategies allows the generation of more consistent and reliable data, helping to improve immunogenicity prediction capabilities in drug development. MHC associated peptide proteomics (MAPPs), Immunogenicity risk assessment, in vitro/ex vivo, biotherapeutics, Major Histocompatibility Complex Class II (MHC II), LC-MS, Immunoaffinity Capture, streptavidin magnetic beads [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
41. Gene therapy in cancer.
- Author
-
Cesur‐Ergün, Büşra and Demir‐Dora, Devrim
- Abstract
Gene therapy, recently frequently investigated, is an alternative treatment method that introduces therapeutic genes into a cancer cell or tissue to cause cell death or slow down the growth of the cancer. This treatment has various strategies such as therapeutic gene activation or silencing of unwanted or defective genes; therefore a wide variety of genes and viral or nonviral vectors are being used in studies. Gene therapy strategies in cancer can be classified as inhibition of oncogene activation, activation of tumor suppressor gene, immunotherapy, suicide gene therapy and antiangiogenic gene therapy. In this review, we explain gene therapy, gene therapy strategies in cancer, approved gene medicines for cancer treatment and future of gene therapy in cancer. Today gene therapy has not yet reached the level of replacing conventional therapies. However, with a better understanding of the mechanism of cancer to determine the right treatment and target, in the future gene therapy, used as monotherapy or in combination with another existing treatment options, is likely to be used as a new medical procedure that will make cancer a controllable disease. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
42. A novel front in sustainable microbial management: computational analysis of curcumin and mangiferin’s synergistic action against Bacillus anthracis
- Author
-
Kahkashan Perveen, Najat A. Bukhari, Najla A. Alshaikh, Suresh Babu Kondaveeti, Jamilah A. Alsulami, Sandip Debnath, and Vinoth Kumarasamy
- Subjects
antibiotics ,biotherapeutics ,mangiferin ,curcumin ,synthetic biology ,molecular docking ,Microbiology ,QR1-502 - Abstract
BackgroundMicroorganisms are crucial in our ecosystem, offering diverse functions and adaptability. The UNGA Science Summit has underscored the importance of understanding microbes in alignment with the UN Sustainable Development Goals. Bacillus anthracis poses significant challenges among various microorganisms due to its harmful effects on both soil and public health. Our study employed computational techniques to investigate the inhibitory effects of curcumin and mangiferin on Bacillus anthracis, with the aim of presenting a novel bio-based approach to microbial management.MethodsEmploying high-throughput screening, we identified potential binding sites on B. anthracis. Molecular docking revealed that curcumin and mangiferin, when synergistically combined, exhibited strong binding affinities at different sites on the bacterium. Our findings demonstrated a significant drop in binding free energy, indicating a stronger interaction when these compounds were used together.FindingsResults of Molecular docking indicated binding energies of −8.45 kcal/mol for mangiferin, −7.68 kcal/mol for curcumin, and a notably higher binding energy of −19.47 kcal/mol for the combination of mangiferin and curcumin with CapD protein. Molecular dynamics simulations further validated these interactions, demonstrating increased stability and structural changes in the bacterium.ConclusionThis study highlights the effectiveness of natural compounds like curcumin and mangiferin in microbial management, especially against challenging pathogens like B. anthracis. It emphasizes the potential of sustainable, nature-based solutions and calls for further empirical research to expand upon these findings.
- Published
- 2024
- Full Text
- View/download PDF
43. Impact of Probiotics on Dairy Production Efficiency
- Author
-
Nalla, Kirankumar, Manda, Naresh Kumar, Dhillon, Harmeet Singh, Kanade, Santosh R, Rokana, Namita, Hess, Matthias, and Puniya, Anil Kumar
- Subjects
Microbiology ,Biological Sciences ,Complementary and Integrative Health ,Nutrition ,Biotechnology ,probiotics ,feed supplements ,biotherapeutics ,dairy production ,gut microbiome ,Environmental Science and Management ,Soil Sciences ,Medical microbiology - Abstract
There has been growing interest on probiotics to enhance weight gain and disease resistance in young calves and to improve the milk yield in lactating animals by reducing the negative energy balance during the peak lactation period. While it has been well established that probiotics modulate the microbial community composition in the gastrointestinal tract, and a probiotic-mediated homeostasis in the rumen could improve feed conversation competence, volatile fatty acid production and nitrogen flow that enhances the milk composition as well as milk production, detailed changes on the molecular and metabolic level prompted by probiotic feed additives are still not understood. Moreover, as living biotherapeutic agents, probiotics have the potential to directly change the gene expression profile of animals by activating the signalling cascade in the host cells. Various direct and indirect components of probiotic approaches to improve the productivity of dairy animals are discussed in this review.
- Published
- 2022
44. Role of Nonclinical Programs in Drug Development
- Author
-
Srivastava, Anup K., Negi, Geeta, Jagadeesh, Gowraganahalli, editor, Balakumar, Pitchai, editor, and Senatore, Fortunato, editor
- Published
- 2023
- Full Text
- View/download PDF
45. Basics of Designing General Toxicology Studies
- Author
-
Peri, Ravikumar, Jagadeesh, Gowraganahalli, editor, Balakumar, Pitchai, editor, and Senatore, Fortunato, editor
- Published
- 2023
- Full Text
- View/download PDF
46. Unveiling Curvularia tuberculata-induced leaf anomalies in Rhododendron ferrugineum: implications in cultural-ecological conservation and harnessing microbial intervention in socio-economic advancement
- Author
-
Juhita Dhar, Aishee Hazra, Riddhisha Patra, Varun Kumar, Vetriselvan Subramaniyan, Vinoth Kumarasamy, Arup Kumar Mitra, Amany A. Sayed, Lotfi Aleya, Fatma M. El-Demerdash, Mikhlid H. Almutairi, Shopnil Akash, Mohamed M. Abdel-Daim, Achal Kant, and Bikram Dhara
- Subjects
plant pathology ,biotherapeutics ,microbe-assisted bioremediation ,synthetic biology ,leaf infection ,R. ferrugineum ,Microbiology ,QR1-502 - Abstract
IntroductionThe research focuses on Rhododendron ferrugineum L., Nepal’s national flower and Uttarakhand’s state tree, thriving in high-altitude mountain ecosystems.Methodology and ResultA study conducted in Himachal Pradesh (Latitude: N 31° 6’ 2.0088”, Longitude: E 77° 10’ 29.9136”) identified leaf anomalies resembling rust-like manifestations in R. ferrugineum. These anomalies were traced back to the pathogenic fungus Curvularia tuberculata, marking the first documented case of its impact on R. ferrugineum in India.DiscussionThis discovery emphasizes the need for vigilant monitoring, disease management research, and conservation efforts to protect the cultural and ecological significance of this iconic shrub. Beyond its immediate findings, the study introduces a novel dimension to Indian flora by associating C. tuberculata with R. ferrugineum, historically linked to monocotyledonous crops. The research methodology combines traditional microscopic examination with advanced genomic sequencing and phylogenetic analysis, enhancing pathogen identification accuracy.Future prospectIn a broader context, this research aligns with the United Nations Sustainable Development Goals (SDGs) by highlighting the importance of environmental preservation, conservation, and sustainable management. It underscores the intricate interplay between biodiversity, cultural heritage, and the need for holistic solutions. Overall, this study calls for proactive measures to protect R. ferrugineum’s cultural and ecological heritage and emphasizes the significance of interdisciplinary approaches in addressing emerging ecological threats.
- Published
- 2024
- Full Text
- View/download PDF
47. Rapid in vitro assessment of the immunogenicity potential of engineered antibody therapeutics through detection of CD4+ T cell interleukin-2 secretion
- Author
-
Yoshiyuki Arata, Shigeki Motoyama, Mariko Yano, Tatsuya Ikuno, Shunsuke Ito, Tomochika Matsushita, Akira Takeiri, Yukari Nishito, Nami Yabuki, Hideaki Mizuno, Zenjiro Sampei, Masayuki Mishima, Masaki Honda, Jumpei Kiyokawa, Hiromi Suzuki, Shuichi Chiba, Mitsuyasu Tabo, and Chiyomi Kubo
- Subjects
Anti-drug antibody ,antibody engineering ,biotherapeutics ,IL-2 ,immunogenicity ,PBMC ,Therapeutics. Pharmacology ,RM1-950 ,Immunologic diseases. Allergy ,RC581-607 - Abstract
ABSTRACTTherapeutic antibodies sometimes elicit anti-drug antibodies (ADAs) that can affect efficacy and safety. Engineered antibodies that contain artificial amino acid sequences are potentially highly immunogenic, but this is currently difficult to predict. Therefore, it is important to efficiently assess immunogenicity during the development of complex antibody-based formats. Here, we present an in vitro peripheral blood mononuclear cell-based assay that can be used to assess immunogenicity potential within 3 days. This method involves examining the frequency and function of interleukin (IL)-2-secreting CD4+ T cells induced by therapeutic antibodies. IL-2-secreting CD4+ T cells seem to be functionally relevant to the immunogenic potential due to their proliferative activity and the expression of several cytokines. The rates of the donors responding to low and high immunogenic proteins, mAb1, and keyhole limpet hemocyanin were 1.3% and 93.5%, respectively. Seven antibodies with known rates of immunogenicity (etanercept, emicizumab, abciximab, romosozumab, blosozumab, humanized anti-human A33 antibody, and bococizumab) induced responses in 1.9%, 3.8%, 6.4%, 10.0%, 29.2%, 43.8%, and 89.5% of donors, respectively. These data are comparable with ADA incidences in clinical settings. Our results show that this assay can contribute to the swift assessment and mechanistic understanding of the immunogenicity of therapeutic antibodies.
- Published
- 2023
- Full Text
- View/download PDF
48. Pushing the frontiers in the fight against antimicrobial resistance: the potential of fecal and maggot therapies
- Author
-
Bashar Haruna Gulumbe and Abdulrakib Abdulrahim
- Subjects
antimicrobial resistance ,biotherapeutics ,fecal microbiota transplantation ,gut microbiome ,maggot debridement therapy ,Medicine ,Medicine (General) ,R5-920 - Abstract
The escalating crisis of antimicrobial resistance (AMR) warrants innovative therapeutic strategies. Fecal microbiota transplantation (FMT) and maggot debridement therapy (MDT) represent paradigm-shifting approaches, leveraging biological systems to mitigate AMR. FMT restores a healthy gut microbiome, providing a biotherapeutic counter to pathogenic bacteria, thereby reducing reliance on traditional antibiotics. Conversely, MDT, a form of bio-debridement, utilizes the antimicrobial secretions of maggots to cleanse wounds and eliminate resistant bacteria. Despite the promise these therapies hold, their broader clinical adoption faces multifaceted challenges including the need for rigorous scientific substantiation, standardized protocols, deepened understanding of mechanisms of action, and surmounting regulatory and public acceptance barriers. However, their potential integration with precision medicine could revolutionize disease management, particularly with antibiotic-resistant infections.
- Published
- 2023
- Full Text
- View/download PDF
49. Microbiota-directed biotherapeutics: considerations for quality and functional assessment
- Author
-
Emily Ef Fekete, Daniel Figeys, and Xu Zhang
- Subjects
Biotherapeutics ,metaproteomics ,microbiome assay ,microbiota ,multi-omics ,Diseases of the digestive system. Gastroenterology ,RC799-869 - Abstract
ABSTRACTMounting evidence points to causative or correlative roles of gut microbiome in the development of a myriad of diseases ranging from gastrointestinal diseases, metabolic diseases to neurological disorders and cancers. Consequently, efforts have been made to develop and apply therapeutics targeting the human microbiome, in particular the gut microbiota, for treating diseases and maintaining wellness. Here we summarize the current development of gut microbiota-directed therapeutics with a focus on novel biotherapeutics, elaborate the need of advanced -omics approaches for evaluating the microbiota-type biotherapeutics, and discuss the clinical and regulatory challenges. We also discuss the development and potential application of ex vivo microbiome assays and in vitro intestinal cellular models in this context. Altogether, this review aims to provide a broad view of promises and challenges of the emerging field of microbiome-directed human healthcare.
- Published
- 2023
- Full Text
- View/download PDF
50. Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies
- Author
-
Maria U. Johansson, Christopher Weinert, Dietrich Alexander Reichardt, Dana Mahler, Dania Diem, Christian Hess, Diana Feusi, Simon Carnal, Julia Tietz, Noreen Giezendanner, Fabio Mario Spiga, David Urech, and Stefan Warmuth
- Subjects
Anti-drug antibodies ,antibody ,antibody fragment ,biotherapeutics ,immunogenicity ,multispecific ,Therapeutics. Pharmacology ,RM1-950 ,Immunologic diseases. Allergy ,RC581-607 - Abstract
ABSTRACTUpon reformatting of an antibody to single-chain variable fragment format, a region in the former variable/constant domain interface of the heavy chain becomes accessible for preexisting (PE) anti-drug antibody (ADA) binding. The region exposed because of this reformatting contains a previously hidden hydrophobic patch. In this study, mutations are introduced in this region to reduce PE ADA reactivity and concomitantly reduce the hydrophobic patch. To enhance our understanding of the importance of individual residues in this region with respect to PE ADA reactivity, a total of 50 molecules for each of two antibodies against different tumor-associated antigens were designed, produced, and characterized by an arsenal of biophysical methods. The aim was to identify suitable mutations that reduce, or completely eliminate, PE ADA reactivity to variable fragments, without compromising biophysical and pharmacodynamic properties. Computational methods were used to pinpoint key residues to mutate and to evaluate designed molecules in silico, in order to reduce the number of molecules to produce and characterize experimentally. Mutation of two threonine residues, Thr101 and Thr146 in the variable heavy domain, proved to be critical to eliminate PE ADA reactivity. This may have important implications in optimizing early drug development for antibody fragment-based therapeutics.
- Published
- 2023
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.