화학공학소재연구정보센터
Journal of Chemical Technology and Biotechnology, Vol.94, No.10, 3344-3355, 2019
A seamless and nonprotein A-based monoclonal antibody purification platform using void-exclusion anion exchange chromatography and electropositive multimodal chromatography enhanced by advance chromatin extraction
BACKGROUND Downstream production of monoclonal antibodies (mAbs) involves several complex unit operations, directly affecting whether they can be used in clinical practice. Protein A affinity chromatography is considered the de facto standard due to its ability to selectively capture antibodies from cell culture supernatant (CCS). Recent studies on various associations of host cell contaminants have shown that downstream purification can be enhanced substantially with advance chromatin removal, facilitating the development of more promising nonaffinity purification platforms. RESULTS In this study, void-exclusion anion exchange chromatography (VEAX) was systematically characterized for its potential use as an initial antibody separation and recovery step. Void exclusion refers to a new chromatographic mode, which is different from the traditional bind-elute and flow-through operations. The new separation method is based on the size of the molecules as well as their surface charges. This process causes the initial immunoglobulin G (IgG) separation to become independent of sample composition (especially for pH and conductivity), eliminating the need to equilibrate the sample. Electropositive multimodal chromatography was then seamlessly incorporated with direct protein loading for further antibody polishing. With the aid of advanced chromatin removal, the two-step purification platform achieved production of an antibody with <1 ppb DNA, This outcome provides a highly promising nonaffinity purification process and a competitive alternative for the current protein A-dominated antibody processing strategy. (c) 2019 Society of Chemical Industry