Improved protein-A chromatography for monoclonal antibody purification.

Quan Chen; Phyllicia Toh; Aina Hoi; Mo Xian; Xinying Peng; Yuansheng Yang; Haibo Zhang; Rui Nian; Wei Zhang

Return

Improved protein-A chromatography for monoclonal antibody purification.

Author: Quan CHEN ¹ ; Phyllicia TOH ² ; Aina HOI ² ; Mo XIAN ¹ ; Xinying PENG ¹ ; Yuansheng YANG ² ; Haibo ZHANG ¹ ; Rui NIAN ¹ ; Wei ZHANG ²
Author Information

1. Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, CAS Key Laboratory of Biobased Materials, Qingdao 266101, Shandong, China.
2. Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Singapore 138668, Singapore.
Publication Type:Journal Article
Keywords: DNA; acidic precipitation; cell culture supernatant; histone; host cell protein; monoclonal antibody; protein A; purification
MeSH: Antibodies, Monoclonal; isolation & purification; Biotechnology; Chromatography, Affinity; DNA; Histones; Hydrogen-Ion Concentration; Immunoglobulin G; isolation & purification; Staphylococcal Protein A; chemistry
From: Chinese Journal of Biotechnology 2016;32(6):807-818
CountryChina
Language:Chinese
Abstract: Therapeutic monoclonal antibodies become the major product class within the biopharmaceutical market. Protein A as the first capture step is still dominant in current platforms for purification of monoclonal antibodies. In this study, we developed a new antibody harvest process that incorporates acidic treatment of cell harvest, demonstrating high process yield, improved clearance of host cell associated contaminants, like non-histone host cell protein, histone, DNA and heteroaggregates. Host protein contamination was reduced about 10-fold compared to protein A loaded with harvest clarified by centrifugation and microfiltration. Turbidity increase of eluted IgG upon pH neutralization was nearly eliminated. Residual levels of impurities in the protein A eluate were achieved that potentially meet requirements of drug substance and thus alleviate the burden for further impurities removal in subsequent chromatography steps. The mechanism of host cell associated contaminants removal during acidic treatment was also explored. After a polishing step by Capto adhere, host cell protein was reduced to less than 5 ppm, DNA less than 1 ppb, histone to undetectable level, heteroaggregates less than 0.01% with total IgG recovery around 87%. This efficient process can be easily integrated into current IgG purification platforms, and may overcome downstream processing challenges.