A Facile Route for Oriented Covalent Immobilization of Recombinant Protein A on Epoxy Agarose Gels: In Situ Generation of Heterofunctional Amino-Epoxy Supports

作者：Yang, Jiamei; Sun, Lifen; Guo, Renling; Yang, Haiyan; Feng, Xiyun; Zhang, Xufeng^*

来源：ChemistrySelect, 2018, 3(37): 10320-10324.

DOI：10.1002/slct.201802256

• Recombinant protein A
• Epoxy agarose
• Oriented immobilization
• Antibodies

The poor reactivity between proteins and epoxy agarose gels limits studies of many protein-agarose conjugates. We present here a novel and simple strategy for oriented covalent immobilization of recombinant protein A (rSpA) on epoxy agarose gels. The method can be basically considered as an immobilization of proteins on the heterofunctional amino-epoxy agarose gels generated in situ. The epoxy agarose gel was directly incubated with rSpA solution containing ethylenediamine (EDA). The presence of EDA during immobilization permits the generation of an in-situ heterofunctional amino-epoxy agarose gel, which permits the ion exchange of rSpA and the following covalent reaction with the epoxy groups. The immobilization efficiency of more than 90% could be achieved by controlling the EDA concentration and pH. Interestingly, the obtained rSpA-agarose conjugates (rSpA@Aga) using the strategy exhibits approximately 20% higher human IgG-binding capacity than those through the immobilization of rSpA on the heterofunctional amino-epoxy agarose gel. Moreover, the resultant rSpA@Aga can selectively adsorb Fc type antibodies and hardly adsorb albumin. Therefore, an effective strategy to develop a high IgG-binding adsorbent is provided, showing potential applications for hemoperfusion in blood purified therapy.

• 出版日期2018-10-9
• 单位云南师范大学