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A Facile Route for Oriented Covalent Immobilization of Recombinant Protein A on Epoxy Agarose Gels: In Situ Generation of Heterofunctional Amino‐Epoxy Supports
Author(s) -
Yang Jiamei,
Sun Lifen,
Guo Renling,
Yang Haiyan,
Feng Xiyun,
Zhang Xufeng
Publication year - 2018
Publication title -
chemistryselect
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.437
H-Index - 34
ISSN - 2365-6549
DOI - 10.1002/slct.201802256
Subject(s) - agarose , epoxy , covalent bond , chemistry , adsorption , chromatography , polymer chemistry , organic chemistry
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.