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A high-throughput purification of monoclonal antibodies from glycoengineered Pichia pastoris
Journal article   Peer reviewed

A high-throughput purification of monoclonal antibodies from glycoengineered Pichia pastoris

Youwei Jiang, Fang Li, Michelle Button, Michael Cukan, Renee Moore, Nathan Sharkey and Huijuan Li
Protein expression and purification, v 74(1), pp 9-15
01 Nov 2010
PMID: 20447459

Abstract

Biochemical Research Methods Biochemistry & Molecular Biology Biotechnology & Applied Microbiology Life Sciences & Biomedicine Science & Technology
Glycoengineered Pichia pastoris provides a unique platform for screening monoclonal antibody (mAb) leads and high expressing strains. A simple, economic, and high-throughput purification for mAb from P. pastoris fermentation has been developed that can be easily operated in various commercially available liquid handlers. The method includes the use of STREAMLINE rProtein A in a 96-well platform and demonstrates good linear alignment and reproducibility in a wide concentration range. The antibody titers measured by the method have less than 15% variation in comparison to spiking titers. The mAb titer and quality obtained from this method are comparable to that from conventional column chromatography. The method can process hundreds of expression screening samples in a day, not only to accurately determine titers, but also to generate milligram quantities of mAb for quality assessment, including purity, folding, glycosylation, and antigen binding affinity. (C) 2010 Elsevier Inc. All rights reserved.

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Web of Science research areas
Biochemical Research Methods
Biochemistry & Molecular Biology
Biotechnology & Applied Microbiology
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