Nian R, Zhang W, Tan L, et al. Advance chromatin extraction improves capture performance of protein A affinity chromatography. J Chromatogr A. 2016;1431:1‐7. doi:10.1016/j.chroma.2015.12.044
Abstract:
Practical effects of advance chromatin removal on performance of protein A affinity chromatography were evaluated using a caprylic acid-allantoin-based extraction method. Lacking this treatment, the practice of increasing loading residence time to increase capacity was shown to increase host protein contamination of the eluted IgG. Advance chromatin extraction suspended that compromise. Protein A ligand leakage from columns loaded with chromatin-extracted harvest was half the level observed on protein A columns loaded with non-extracted harvest. Columns loaded with chromatin-extracted harvest were cleaned more effectively by 50-100mM NaOH than columns loaded with non-extracted harvest that were cleaned with 250-500mM NaOH. Two protein A media with IgG capacities in excess of 50g/L were loaded with chromatin-extracted harvest, washed with 2.0M NaCl before elution, and the eluted IgG fraction titrated to pH 5.5 before microfiltration. Host protein contamination in the filtrate was reduced to
License type:
http://creativecommons.org/licenses/by-nc-nd/4.0/
Funding Info:
This research / project is supported by the Exploit Technologies Pte Ltd, A*STAR, under its ETPL GAP-Funded (Project No. ETPL/12-R15GAP-0009).