Advance chromatin extraction enhances performance and productivity of cation exchange chromatography based capture of Immunoglobulin G monoclonal antibodies.
The impact of host cell derived chromatin was investigated on the performance and productivity of cation exchange chromatography as a method for capture purification of an IgG monoclonal antibody. Cell culture supernatant was prepared for loading by titration to pH 6.0, dilution with water to a conductivity of 4mS/cm, then microfiltration to remove solids. DNA content was reduced 99% to 30ppm, histone host cell protein content by 76% to 6300ppm, non histone host cell protein content by 15% to 321,000ppm, and aggregates from 33% to 15% IgG recovery was 83% An alternative preparation was performed, adding octanoic acid, allantoin, and electropositive particles to the harvest at pH 5.3, then removing solids. DNA content was reduced to<1 ppb, histones became undetectable, non histones were reduced to 24,000ppm, and aggregates were reduced to 2.4% IgG recovery was 95% This treatment increased dynamic capacity (DBC) of cation exchange capture to 173g/L and enabled the column to reduce non histone host proteins to 671ppm. Step recovery was 99% A single multimodal polishing step further reduced them to 15ppm and aggregates to <0.1% Overall process recovery was 89% Productivity at feed stream IgG concentrations of 5 10g/L was roughly double the productivity of a same size protein A column with a DBC of 55g/L.
Journal of chromatography. A