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Integrated solution to purification challenges in the manufacture of a soluble recombinant protein in E. coli

✍ Scribed by Maire H. Caparon; Kevin J. Rust; Alan K. Hunter; Joseph K. McLaughlin; Kristen E. Thomas; John T. Herberg; Robert E. Shell; Paul B. Lanter; Bruce F. Bishop; Robert L. Dufield; Xing Wang; Sa V. Ho

Publisher
John Wiley and Sons
Year
2010
Tongue
English
Weight
851 KB
Volume
105
Category
Article
ISSN
0006-3592

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✦ Synopsis

Abstract

Apolipoprotein A 1 Milano (ApoA‐1M), the protein component of a high‐density lipoprotein (HDL) mimic with promising potential for reduction of atherosclerotic plaque, is produced at large scale by expression in E. coli. Significant difficulty with clearance of host cell proteins (HCPs) was experienced in the original manufacturing process despite a lengthy downstream purification train. Analysis of purified protein solutions and intermediate process samples led to identification of several major HCPs co‐purifying with the product and a bacterial protease potentially causing a specific truncation of ApoA‐1M found in the final product. Deletion of these genes from the original host strain succeeded in substantially reducing the levels of HCPs and the truncated species without adversely affecting the overall fermentation productivity, contributing to a much more efficient and robust new manufacturing process. Biotechnol. Bioeng. 2010; 105: 239–249. © 2009 Wiley Periodicals, Inc.

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