ABHINAV SHUKLA PROTEASE PDF

Host cell protein HCP contaminant clearance is a significant concern during downstream process development for biopharmaceuticals. Protein A chromatography as a capture step for monoclonal antibodies and Fc fusion proteins can clear a large proportion of these impurities from cell culture harvest. Nevertheless, remaining levels of this process-related impurity class do present significant constraints on the rapid development of effective and robust polishing steps. Conventionally, an intermediate pH wash is employed between column loading and elution to minimize HCP levels after Protein A chromatography. A significant mechanistic finding presented in this work is that HCP contaminants that persist following Protein A capture predominantly comprise species that associate with the product in preference to direct interaction with the chromatographic resin. A higher wash pH to preserve product--Protein A binding along with the use of additive combinations to disrupt interactions between HCPs and the product are investigated.

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Host cell protein HCP contaminant clearance is a significant concern during downstream process development for biopharmaceuticals.

Protein A chromatography as a capture step for monoclonal antibodies and Fc fusion proteins can clear a large proportion of these impurities from cell culture harvest. Nevertheless, remaining levels of this process-related impurity class do present significant constraints on the rapid development of effective and robust polishing steps.

Conventionally, an intermediate pH wash is employed between column loading and elution to minimize HCP levels after Protein A chromatography. A significant mechanistic finding presented in this work is that HCP contaminants that persist following Protein A capture predominantly comprise species that associate with the product in preference to direct interaction with the chromatographic resin.

A higher wash pH to preserve product--Protein A binding along with the use of additive combinations to disrupt interactions between HCPs and the product are investigated.

This strategy was successfully applied to develop a broadly applicable wash condition that has the potential for eliminating the need for product specific optimization of wash conditions. Use of this and other similar wash conditions are anticipated to aid the rapid development of effective downstream processes for monoclonal antibodies and Fc fusion proteins resulting in their rapid introduction into clinical trials.

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Similar articles Demonstration of robust host cell protein clearance in biopharmaceutical downstream processes. Shukla AA, et al. Biotechnol Prog. Epub Apr PMID: Purification of an Fc-fusion biologic: clearance of multiple product related impurities by hydrophobic interaction chromatography.

Evans DR, et al. J Chromatogr A. Epub Jul Protein aggregation kinetics during Protein A chromatography. Case study for an Fc fusion protein. Epub Sep Downstream processing of monoclonal antibodies--application of platform approaches. Epub Oct PMID: Review. Effective strategies for host cell protein clearance in downstream processing of monoclonal antibodies and Fc-fusion proteins.

Protein Expr Purif. Show more similar articles See all similar articles. Cited by 24 articles Modelling of pH-dependence to develop a strategy for stabilising mAbs at acidic steps in production. Hebditch M, et al. Comput Struct Biotechnol J. Enhancing Protein A performance in mAb processing: A method to reduce and rapidly evaluate host cell DNA levels during primary clarification.

Koehler KC, et al. Epub Aug Lavoie RA, et al. Int J Mol Sci. Luo H, et al. Epub Nov 4. Application of enhanced electronegative multimodal chromatography as the primary capture step for immunoglobulin G purification. Wang Y, et al. AMB Express. Show more "Cited by" articles See all "Cited by" articles. Substances Antibodies, Monoclonal Actions. Immunoglobulin Fc Fragments Actions. Recombinant Fusion Proteins Actions. Staphylococcal Protein A Actions. Full-text links [x] Wiley. Copy Download.

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Abhinav Shukla

Abhinav Shukla is an Indian model and television and film actor. He has acted in many TV shows. Abhinav was born in Ludhiana , Punjab. His father, Dr. He started his career in the Indian television industry with Jersey No.

LA CIENCIA DE LA ORACION YIYE AVILA PDF

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