1. Biocatalytic Method for Producing an Affinity Resin for the Isolation of Immunoglobulins.
- Author
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Tereshin, Mikhail N., Melikhova, Tatiana D., Eletskaya, Barbara Z., Ivanova, Elena A., Onoprienko, Lyudmila V., Makarov, Dmitry A., Razumikhin, Mikhail V., Myagkikh, Igor V., Fabrichniy, Igor P., and Stepanenko, Vasiliy N.
- Subjects
MUTANT proteins ,AFFINITY chromatography ,AMINO group ,IMMUNOGLOBULINS ,MONOCLONAL antibodies - Abstract
Affinity chromatography is a widely used technique for antibody isolation. This article presents the successful synthesis of a novel affinity resin with a mutant form of protein A (BsrtA) immobilized on it as a ligand. The key aspect of the described process is the biocatalytic immobilization of the ligand onto the matrix using the sortase A enzyme. Moreover, we used a matrix with primary amino groups without modification, which greatly simplifies the synthesis process. The resulting resin shows a high dynamic binding capacity (up to 50 mg IgG per 1 mL of sorbent). It also demonstrates high tolerance to 0.1 M NaOH treatment and maintains its effectiveness even after 100 binding, elution, and sanitization cycles. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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