Non-immunospecific association of immunoglobulin G with chromatin during elution from protein A inflates host contamination, aggregate content, and antibody loss
| Autor: | Yuansheng Yang, Qiaohui Yang, Rui Nian, Pete Gagnon, Chiew Ling Lim |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
IgG
Microfiltration Clinical Biochemistry Protein aggregation Buffers Biochemistry Immunoglobulin G Neutralization Chromatography Affinity Analytical Chemistry Protein Aggregates Host contamination Aggregation Recovery Staphylococcal Protein A Chromatography biology Chemistry Elution Organic Chemistry Antibodies Monoclonal General Medicine Hydrogen-Ion Concentration Chromatin biology.protein Molecular Medicine Protein A Antibody |
| Zdroj: | Journal of Chromatography A. :151-160 |
| ISSN: | 0021-9673 |
| DOI: | 10.1016/j.chroma.2015.07.017 |
| Popis: | 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 (100mM 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. |
| Databáze: | OpenAIRE |
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