00558nas a2200109   4500008004100000245009500041210006900136260009100205100002000296700002000316856011200336       2017                        eng d00aImplementation of QbD for development of a downstream process for a therapeutic biosimilar0 aImplementation of QbD for development of a downstream process fo  aSan Francisco, CAbAMER CHEMICAL SOC, 1155 16TH ST, NW, WASHINGTON, DC 20036 USAcAPR 1 aRathore, Anurag1 aBhambure, Rahul  uhttps://library.ncl.res.in/content/implementation-qbd-development-downstream-process-therapeutic-biosimilar02022nas a2200253   4500008004100000022001400041245010800055210006900163260000900232300001100241490000700252520118300259653002301442653002901465653001501494653002601509653001401535100002001549700001701569700001901586700002001605700002001625856012301645       2020                        eng d  a0734-975000aEngineering staphylococcal protein A for high-throughput affinity purification of monoclonal antibodies0 aEngineering staphylococcal protein A for highthroughput affinity  cNOV   a1076320 v443 a<p>Protein A chromatography is one of the most widely used purification steps in the manufacturing of the various classes of recombinant and non-recombinant antibodies. Due to the higher cost, lower binding capacity, and limited life cycle of Protein A ligand, this affinity-based purification step is often one of the most significant contributors to the cost of manufacturing of monoclonal antibody (mAb) products. In the last decade, there has been significant progress in improving the Protein A chromatography throughput by designing new engineered Staphylococcal Protein A (SPA) variants with higher dynamic binding capacity, considerable alkaline tolerance, and mild acidic elution pH. This review aims at summarizing the various protein engineering approaches used for improving the throughput of the Protein A-based affinity purification of various immunoglobulins. With biopharmaceutical producers operating under ever-increasing pressure towards reducing the cost of manufacturing, these advances in engineered protein A variants will help in processing larger cell culture volumes with high throughput and thereby significantly lower the cost of raw materials.</p>
10aAlkaline tolerance10aDynamic binding capacity10aElution pH10aMonoclonal antibodies10aProtein A1 aAmritkar, Vinod1 aAdat, Satish1 aTejwani, Vijay1 aRathore, Anurag1 aBhambure, Rahul  uhttps://library.ncl.res.in/content/engineering-staphylococcal-protein-high-throughput-affinity-purification-monoclonal02178nas a2200217   4500008004100000245011100041210006900152260000800221300001200229490000800241520143400249100001801683700001501701700002001716700001801736700002101754700001701775700002001792700002001812856012801832       2022                        eng d00aDimerization of SARS-CoV-2 nucleocapsid protein affects sensitivity of ELISA based diagnostics of COVID-190 aDimerization of SARSCoV2 nucleocapsid protein affects sensitivit  cMAR  a428-4370 v2003 a<p>Nucleocapsid protein (N protein) is the primary antigen of the virus for development of sensitive diagnostic assays of COVID-19. In this paper, we demonstrate the significant impact of dimerization of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) N-protein on sensitivity of enzyme-linked immunosorbent assay (ELISA) based diagnostics. The expressed purified protein from E. coli is composed of dimeric and monomeric forms, which have been further characterized using biophysical and immunological techniques. Indirect ELISA indicated elevated susceptibility of the dimeric form of the nucleocapsid protein for identification of protein-specific monoclonal antibody as compared to the monomeric form. This finding also confirmed with the modelled structure of monomeric and dimeric nucleocapsid protein via HHPred software and its solvent accessible surface area, which indicates higher stability and antigenicity of the dimeric type as compared to the monomeric form. The sensitivity and specificity of the ELISA at 95% CI are 99.0% (94.5-99.9) and 95.0% (83.0-99.4), respectively, for the highest purified dimeric form of the N protein. As a result, using the highest purified dimeric form will improve the sensitivity of the current nucleocapsid-dependent ELISA for COVID-19 diagnosis, and manufacturers should monitor and maintain the monomer-dimer composition for accurate and robust diagnostics.</p>
1 aKhan, Wajihul1 aKhan, Nida1 aMishra, Avinash1 aGupta, Surbhi1 aBansode, Vikrant1 aMehta, Deepa1 aBhambure, Rahul1 aRathore, Anurag  uhttps://library.ncl.res.in/content/dimerization-sars-cov-2-nucleocapsid-protein-affects-sensitivity-elisa-based-diagnostics