Optimizing selectivity of anion hydrophobic multimodal chromatography for purification of a single-chain variable fragment.

ABSTRACT: Single-chain variable fragments (scFv) are widely used in several fields. However, they can be challenging to purify unless using expensive Protein L-based affinity adsorbents or affinity tags. In this work, a purification process for a scFv using mixed-mode (MM) chromatography was developed by design of experiments (DoE) and proteomics for host cell protein (HCP) quantification. Capture of scFv from human embryonic kidney 293 (HEK293) cell feedstocks was performed by hydrophobic charge induction chromatography (MEP HyperCel™), whereafter polishing was performed by anion hydrophobic MM chromatography (Capto Adhere™). The DoE designs of the polishing step included both binding and flow-through modes, the latter being the standard mode for HCP removal. Chromatography with Capto Adhere™ in bi

SUBMITTER: Sakhnini LI 

PROVIDER: S-EPMC6999250 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature