Journal of Chromatography & Separation Techniques
Open Access
ISSN: 2157-7064
+44 1300 500008
ISSN: 2157-7064
+44 1300 500008
Dong-Qiang Lin and Shan-Jing Yao
Zhejiang University, China
Posters & Accepted Abstracts: J Chromatogr Sep Tech
For biopharmaceuticals downstream processes, it always requires highly productive and robust technologies to improve the process efficiency. Expanded-bed adsorption (EBA) is an innovative technology that allows capturing proteins directly from un-clarified feedstock, such as cell culture broth and homogenization. EBA technology combines solid-liquid separation with an adsorption step in a single-unit operation, aiming at increased overall yield, reduced operational time, and less requirements for capital investment and consumables. Mixed-mode chromatography (MMC) is a novel technology for bio-product separation, which combines multiple binding modes like hydrophobic and electrostatic interactions, hydrogen bonding, etc. High capacity, salt-tolerance, good selectivity and relatively low cost are the major advantages of MMC for direct capture process. In the present work two chromatographic techniques, EBA and MMC, were integrated to develop new separation technology, mixed-mode EBA, improving the protein capture efficiency and reducing the pretreatments on the feedstock, such as clarification, dilution and salt-adjustment. Several MMC ligands were coupled onto typical matrices (densified agarose beads) for EBA. The static adsorption, adsorption kinetics and dynamic binding were investigated, and the effects of pH and salt addition were evaluated. New technology was challenged with two typical biopharmaceutical processes, monoclonal antibody (mAb) capture from CHO cell culture broth and recombinant human albumin serum (rHSA) isolation from Pichia pastoris fermentation broth. After the optimization of operation conditions, high separation efficiency (purity, recovery, productivity) was obtained. The results demonstrated that mixed-mode EBA, combining the advantages of EBA and MMC, would be a promising new platform for protein capture with reduced feedstock pretreatments, high efficiency and relative low cost. New technology developed in the present work could also be expanded to other bio-product processes.
Email: lindq@zju.edu.cn