Search

Characterization and quantification of virus-like particles (VLPs) through high performance liquid chromatography (HPLC)-based methods are challenging because of their large size, structural complexity, internal structural heterogeneity, and instability. Analytical techniques are essential to monitor morphology and internal structural heterogeneity at each process stage. Common analytical tools used in VLP characterization are microscopic techniques (such as transmission electron microscopy [TEM], atomic force-field microscopy [AFM], cryo-electron microscope [cryo-EM]), biochemical techniques (SDS-PAGE, western blotting), and light scattering techniques (such as dynamic light scattering [DLS], nanoparticle tracking analysis [NTA], and size-exclusion chromatography coupled with multi-angle light scattering [SEC-MALS]). However, these techniques are semi-quantitative and do not address morphology and internal heterogeneity. Therefore, HPLC-based techniques are sensitive, robust, and offer better resolution. The purity and titer of VLPs at any process stage can be monitored by reversed-phase chromatography and morphology, and stability-related issues can be monitored by the combination of HPLC and light scattering techniques like SEC-MALS. Challenges in HPLC-based methods are choosing columns with the right pore size and surface chemistry and effective sample preparations, as VLPs are very unstable and prone to fragmentation at process stages and the low titre of the VLPs. This article discusses the challenges and effective solutions for HPLC-based analytical characterization of VLPs., VIRUS-LIKE PARTICLES (VLPS) ARE nanoparticle-sized, multimeric, self-assembled protein complexes. They are non-infectious, as they lack the genetic material of parental viruses (1). VLPs trigger both cell-mediated and humoral-mediated immune responses, [...]

Biopharmaceutical analysis is a rapidly evolving field that requires the development of new technologies and methods to keep pace with the increasing complexity of biologics. One of the most promising areas of research is the use of single-cell omics and microfluidic chips for the analysis of biopharmaceuticals. Single-cell omics has revolutionized our understanding of cellular heterogeneity, while microfluidic chips have enabled high-throughput analysis of single cells that provide an understanding of the complex biological network that complements the genomics and transcriptomics studies. This article will explore some of the emerging trends and technologies in biopharmaceutical analysis, with a particular focus on single-cell omics and microfluidic chips. We will also discuss the developments in ambient ionization mass spectrometry such as sub nanoampere ionization and the potential of low current ionization in studying cell-to-cell heterogeneity and its role in metabolomics., VARIATIONS DISPLAYED by the cell populations in their DNA, RNA, proteins, and metabolites lead to significant heterogeneity and differences in cell states (1). An understanding of the cell-to-cell variations can [...]

Gene therapy is a procedure that enables gene improvement by means of the correction of the altered (mutated) genes, or targets site-specific modifications that have therapeutic treatment as a target [...]

Ion pairing agents are frequently used in liquid chromatography (LC), especially in reversed-phase liquid chromatography (RPLC), to increase the separation and retention of charged analytes. However, their use in other modes of HPLC, such as sizeexclusion HPLC (SE-HPLC) and ion exchange chromatography (IEC) has not been extensively explored. In RPLC, the target analytes may have charged functional groups, which makes it difficult to maintain them on the hydrophobic stationary phase. This difficulty is often overcome by the inclusion of an ion pairing agent, which introduces ionic contacts and strengthens the hydrophobic connections between the analytes and stationary phase. The separation mechanisms in both SE-HPLC and IEC rely on size and charge differences, respectively, between analytes. Thus, they are naturally constructed to separate analytes without the requiring extra ion pairing agents. In this study, we introduced an ion pairing agent (, Reversed-phase liquid chromatography (RPLC) is a simple and user-friendly technology that is often used for separation of bioanalytes, thereby accounting for up to 70-80% of bioseparations (1). Most acidic samples [...]

Over the last several years, our definition of biotherapeutics has evolved from solely protein-based therapeutics, such as monoclonal antibodies (mAbs), to include such product classes as cell and gene therapies. Cell and gene therapies are unique products that require innovative and new approaches to formulation, specifically related to drug delivery. One such delivery modality that has gained significant traction is lipid nanoparticles (LNPs), which, as their name suggests, are nanoparticles comprised of lipids. The first LNP product approved by the United States Food and Drug Administration, was the liposome-encapsulated Doxil, whereas, in 2018, the United States Food and Drug Administration approved the small interfering RNA (siRNA) product Onpattro, using lipid nanoparticles as part of its formulation. Since then, LNPs have gained more international attention in their role in drug delivery because of the mRNA-based COVID-19 vaccines. LNPs in the biopharmaceutical space are still in their infancy related to our understanding of them and how to characterize them, presenting both challenges and opportunities. In this column, we discuss some of those challenges and opportunities and look at the current and future landscape of the most common analytical tools used to analyze and characterize LNP, As our understanding of biotherapeutics expands to include new products such as cell and gene therapies, we are presented with new challenges and opportunities in the characterization of these products. [...]

Multi-attribute methods (MAMs) are becoming increasingly popular for their ability to analyze multiple critical quality attributes (CQA) in a single workflow. This capability becomes particularly attractive for a product class [...]

Analytical methods are used in the biopharmaceutical industry to ensure the quality, efficacy, and safety of drug substances and drug products. One of the fundamental measures of the quality of a drug substance or drug product, including biopharmaceuticals, is the stability of the active pharmaceutical ingredient (API). In fact, the first International Council for Harmonization (ICH) guideline on quality, ICH Q1, is for drug stability. In this column, we look at drug stability in general, differences between largeand small-molecule stability (ICH Q1 and ICH Q5), as well as the analytical methods used to measure the stability of a product. Although there are several analytical methodologies that can be used, and we touch on those briefly, we focus mostly on the chromatography and mass spectrometry methods used to characterize drug stability., At the center of everything we do in the biopharmaceutical industry should be patients and ensuring that their medicines are high quality, safe, and effective. A cornerstone of any drug [...]

The landscape of biopharmaceutical analysis features different analytical requirements in terms of instrumentation and type of analysis (QC, RTRT, and in-process). One of the questions that often crops up for routine analysis is 'How much replication is enough?' Should all the samples be run in triplicate irrespective of the type of analysis, or does the type of analysis (such as charge or size variants) have any bearing on the number of optimal replicates?, There are various factors that the user needs to consider in the matter of biopharmaceutical analysis--first, the objective of replication, and second, the trade-off with respect to cost and time. [...]