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Tag-assisted protein purification is a method of choice for both academic researches and large-scale industrial demands. Application of the purification tags in the protein production process can help to save time and cost, but the design and application of tagged fusion proteins are challenging. An appropriate tagging strategy must provide sufficient expression yield and high purity for the final protein products while preserving their native structure and function. Thanks to the recent advances in the bioinformatics and emergence of high-throughput techniques (e.g. SEREX), many new tags are introduced to the market. A variety of interfering and non-interfering tags have currently broadened their application scope beyond the traditional use as a simple purification tool. They can take part in many biochemical and analytical features and act as solubility and protein expression enhancers, probe tracker for online visualization, detectors of post-translational modifications, and carrier-driven tags. Given the variability and growing number of the purification tags, here we reviewed the protein- and peptide-structured purification tags used in the affinity, ion-exchange, reverse phase, and immobilized metal ion affinity chromatographies. We highlighted the demand for purification tags in the pharmaceutical industry and discussed the impact of self-cleavable tags, aggregating tags, and nanotechnology on both the column-based and column-free purification techniques. • The application of tags for protein purification will reduce the cost of preparing the chromatographic columns. • A suitable tagging system may provide a sufficient expression yield and high purity of the protein product. • The introduction of the self-cleavable tags and aggregating tags have been influential in the protein purification industry. • The application of the aggregating tags in a column-free manner can facilitate higher protein expression yield. • Emerging technologies, including bioinformatics, SELEX, and nanotechnology, have helped to improve technical limitations. [ABSTRACT FROM AUTHOR]

Evaluating the effectiveness of drugs on an individual basis before prescribing them has an influential role in choosing a successful treatment plan. The advent of microfluidic devices has provided us with more real platforms capable of mimicking the natural microenvironment (scales, shear stress, diffusion transport, etc.). This research has introduced a truncated microfluidic gradient maker that can create a linear concentration gradient of trastuzumab (showing good correlation (r(9) = 0.9968) with simulation data) with a wide range of flow rates (20–190 µl/min) and investigate its effect on SK-BR-3 spheroid cells. The results of trastuzumab efficacy on 3D spheroid cells in the microchip were compared with the conventional 2D cell cultured by MTT assay at 24 h, 48 h, and 72 h. The results showed that trastuzumab had dose-dependent effects in both 2D and spheroid cell cultures. However, the IC50 values for trastuzumab were significantly higher in 2D cell culture (560 ± 23 µg/mL) compared to 3D spheroid models (407 ± 17 µg/mL). Moreover, compared with conventional drug sensitivity tests, this microchip required fewer samples and reagents and a lower risk of fluctuation and contamination due to automatic medium exchange. [Display omitted] • A novel truncated microfluidic chip which can generate nine different dosages of the drug in a smaller design was constructed. • Effects of the different dosages of trastuzumab on SK-BR-3 spheroids were observed. • The performance of trastuzumab on SK-BR-3 spheroids cultured in chip and 2d conventional model was compared. [ABSTRACT FROM AUTHOR]

• MERS-CoV, SARS-CoV-1/2 as members of β-CoVs family have many similar pathogenicity. • Frequency of emerging β-CoVs infections reported in males are higher than females. • Sex-hormones and X-chromosome related genes may be the cause of these sex differences. • Immune response of women might protect further against MERS, SARS-CoV1/2 infections. • Sex-based immune-responses may provide new insights for upcoming immunotherapies. Emerging beta-coronaviruses (β-CoVs), including Severe Acute Respiratory Syndrome CoV-1 (SARS-CoV-1), Middle East Respiratory Syndrome-CoV (MERS-CoV), and Severe Acute Respiratory Syndrome CoV-2 (SARS-CoV-2, the cause of COVID19) are responsible for acute respiratory illnesses in human. The epidemiological features of the SARS, MERS, and new COVID-19 have revealed sex-dependent variations in the infection, frequency, treatment, and fatality rates of these syndromes. Females are likely less susceptible to viral infections, perhaps due to their steroid hormone levels, the impact of X-linked genes, and the sex-based immune responses. Although mostly inactive, the X chromosome makes the female's immune system more robust. The extra immune-regulatory genes of the X chromosome are associated with lower levels of viral load and decreased infection rate. Moreover, a higher titer of the antibodies and their longer blood circulation half-life are involved in a more durable immune protection in females. The activation rate of the immune cells and the production of TLR7 and IFN are more prominent in females. Although the bi-allelic expression of the immune regulatory genes can sometimes lead to autoimmune reactions, the higher titer of TLR7 in females is further associated with a stronger anti-viral immune response. Considering these sex-related differences and the similarities between the SARS, MERS, and COVID-19, we will discuss them in immune responses against the β-CoVs-associated syndromes. We aim to provide information on sex-based disease susceptibility and response. A better understanding of the evasion strategies of pathogens and the host immune responses can provide worthful insights into immunotherapy, and vaccine development approaches. [ABSTRACT FROM AUTHOR]