International Journal of Molecular Sciences (Jan 2024)

Affinity Resins for the Isolation of Immunoglobulins G Obtained Using Biocatalytic Technology

  • Mikhail N. Tereshin,
  • Tatiana D. Melikhova,
  • Barbara Z. Eletskaya,
  • Olga B. Ksenofontova,
  • Pavel V. Pantyushenko,
  • Maria Ya. Berzina,
  • Igor Ivanov,
  • Igor V. Myagkikh,
  • Vasiliy N. Stepanenko

DOI
https://doi.org/10.3390/ijms25031367
Journal volume & issue
Vol. 25, no. 3
p. 1367

Abstract

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Affinity chromatography resins that are obtained by conjugation of matrices with proteins of bacterial origin, like protein A, are frequently used for the purification of numerous therapeutic monoclonal antibodies. This article presents the development of a biocatalytic method for the production of novel affinity resins with an immobilized mutant form of protein A via sortase A mediated reaction. The conditions for activation of the agarose Seplife 6FF matrix, selection of different types of linkers with free amino groups and conditions for immobilization of recombinant protein A on the surface of the activated matrix were studied. Finally, the basic operational properties, like dynamic binding capacity (DBC), temperature dependance of DBC and stability during the cleaning-in-place process of the affinity resin with the Gly-Gly-EDA-Gly-Gly linker, were assessed using recombinant hyperchimeric monoclonal antibodies. The main characteristics show comparable results with the widely used commercial samples.

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