Combined protein A imprinting and cryogelation for production of spherical affinity material


ASLIYÜCE ÇOBAN S., Mattiasson B., DENİZLİ A.

BIOMEDICAL CHROMATOGRAPHY, vol.33, no.10, 2019 (Peer-Reviewed Journal) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 33 Issue: 10
  • Publication Date: 2019
  • Doi Number: 10.1002/bmc.4605
  • Journal Name: BIOMEDICAL CHROMATOGRAPHY
  • Journal Indexes: Science Citation Index Expanded, Scopus

Abstract

Cryogels have been demonstrated to be efficient when applied for protein isolation. Owing to their macroporous structure, cryogels can also be used for treating particle-containing material, e.g. cell homogenates. Another challenging development in protein purification technology is the use of molecularly imprinted polymers (MIPs). These MIPs are robust and can be used repeatedly. The paper presents a new technology that combine the formation of cryogel beads concomitantly with making imprints of a protein. Protein A was chosen as the print molecule which was also be the target in the purification step. The present paper describes a new method to produce protein-imprinted cryogel beads. The protein-imprinted material was characterized and the separation properties were evaluated with regard to both the target protein and whole cells with target protein exposed on the cell surface. The maximum protein A adsorption was 18.1 mg/g of wet cryogel beads. The selectivity coefficient of protein A-imprinted cryogel beads for protein A was 5.44 and 12.56 times greater than for the Fc fragment of IgG and protein G, respectively.