Improved Performance of Recombinant Protein A Immobilized on Agarose Beads by Site-Specific Conjugation.

ABSTRACT: Protein A affinity adsorbent with high antibody-binding capacity plays a prominent part in the purification of biopharmaceuticals to decrease the manufacturing costs. We describe a site-specific covalent conjugation strategy for protein A to immobilize on agarose beads. Recombinant protein A, which has one cysteine introduced at the C terminus through genetic engineering technology, was immobilized site-specifically on maleimide-functionalized agarose beads by the thiol-maleimide reaction. As a comparison, the recombinant protein A was randomly immobilized on the aldehyde-functionalized agarose beads via free amino groups on the protein surface. The site-specific conjugation of recombinant protein A on the agarose beads was validated through the assay of free SH groups on the adsorbents us

SUBMITTER: Zhang X 

PROVIDER: S-EPMC6044777 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature