Purification of antibodies by precipitating impurities using Polyethylene Glycol to enable a two chromatography step process
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- 作者：Glen Giese ; Adam Myrold ; Jeffrey Gorrell ; Josefine Persson
- 关键词：AIEX ; anion exchange ; CIEX ; cation exchange ; CV ; column volume ; DOE ; Design of Experiments ; LRV ; Log Reduction Value ; HCP ; host cell protein ; HMWS ; high molecular weight species ; HPLC ; High Performance Liquid Chromatography ; MAb ; monoclonal antibody ; MMV
- 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
- 出版年：2013
- 出版时间：1 November, 2013
- 年：2013
- 卷：938
- 期：Complete
- 页码：14-21
- 全文大小：599 K
文摘
The purification of antibodies by precipitating impurities using Polyethylene Glycol (PEG) was assessed with the objective of developing a two chromatography column purification process. A PEG precipitation method was evaluated for use in the industrial purification of recombinant monoclonal antibodies (MAbs). Effective and robust precipitation conditions including PEG concentration, pH, temperature, time, and protein concentration were identified for several different MAbs. A recovery process using two chromatography steps in combination with PEG precipitation gave acceptable yield and purity levels for IgG1 and IgG4 antibodies with a broad range of isoelectric points (pI). PEG precipitation removed host cell proteins (HCPs), high molecular weight species (HMWS), leached Protein A ligand, and host cell DNA to acceptable levels when run under appropriate conditions, and some endogenous virus removal was achieved.