期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1408, 期 -, 页码 151-160出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2015.07.017
关键词
Protein A; IgG; Chromatin; Host contamination; Aggregation; Recovery
资金
- Exploit Technologies Pte. Ltd.
- Validated Biosystems
- Biomedical Research Council of the Singapore Agency for Science and Technology Research
Monoclonal IgG at pH 3.5 expressed a tendency to self-associate and associate non-specifically with surfaces, including the surfaces of precipitated chromatin heteroaggregates. The tendency was elevated with protein A-eluted IgG still in elution buffer (100 mM acetate, pH 3.5). Association of IgG with chromatin elements under protein A elution conditions amplified host protein contamination of the elution fraction about 15-fold, caused formation of aggregates that persisted after pH neutralization, and imposed an approximate 5% loss on IgG recovery. Neutralization released eluted IgG from its low pH associations with chromatin and caused heteroaggregate remnants to associate into large particles easily removed by microfiltration. Most effective host contaminant clearance was achieved by filtration after neutralization to pH 5.5. All chromatin-mediated liabilities were suspended by extraction of chromatin heteroaggregates in advance of protein A. (C) 2015 The Authors. Published by Elsevier B.V.
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