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Article

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

by
Mikhail N. Tereshin
1,2,
Tatiana D. Melikhova
1,
Barbara Z. Eletskaya
1,
Olga B. Ksenofontova
1,
Pavel V. Pantyushenko
1,
Maria Ya. Berzina
1,*,
Igor Ivanov
2,
Igor V. Myagkikh
1 and
Vasiliy N. Stepanenko
1,2
1
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya St. 16/10, 117437 Moscow, Russia
2
Lomonosov Institute of Fine Chemical Technologies, MIREA—Russian Technological University, Vernadskogo pr. 86, 119571 Moscow, Russia
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2024, 25(3), 1367; https://doi.org/10.3390/ijms25031367
Submission received: 12 December 2023 / Revised: 15 January 2024 / Accepted: 19 January 2024 / Published: 23 January 2024
(This article belongs to the Section Biochemistry)
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Abstract

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.
Keywords: protein A chromatography; monoclonal antibodies; sortase A protein A chromatography; monoclonal antibodies; sortase A

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MDPI and ACS Style

Tereshin, M.N.; Melikhova, T.D.; Eletskaya, B.Z.; Ksenofontova, O.B.; Pantyushenko, P.V.; Berzina, M.Y.; Ivanov, I.; Myagkikh, I.V.; Stepanenko, V.N. Affinity Resins for the Isolation of Immunoglobulins G Obtained Using Biocatalytic Technology. Int. J. Mol. Sci. 2024, 25, 1367. https://doi.org/10.3390/ijms25031367

AMA Style

Tereshin MN, Melikhova TD, Eletskaya BZ, Ksenofontova OB, Pantyushenko PV, Berzina MY, Ivanov I, Myagkikh IV, Stepanenko VN. Affinity Resins for the Isolation of Immunoglobulins G Obtained Using Biocatalytic Technology. International Journal of Molecular Sciences. 2024; 25(3):1367. https://doi.org/10.3390/ijms25031367

Chicago/Turabian Style

Tereshin, Mikhail N., Tatiana D. Melikhova, Barbara Z. Eletskaya, Olga B. Ksenofontova, Pavel V. Pantyushenko, Maria Ya. Berzina, Igor Ivanov, Igor V. Myagkikh, and Vasiliy N. Stepanenko. 2024. "Affinity Resins for the Isolation of Immunoglobulins G Obtained Using Biocatalytic Technology" International Journal of Molecular Sciences 25, no. 3: 1367. https://doi.org/10.3390/ijms25031367

APA Style

Tereshin, M. N., Melikhova, T. D., Eletskaya, B. Z., Ksenofontova, O. B., Pantyushenko, P. V., Berzina, M. Y., Ivanov, I., Myagkikh, I. V., & Stepanenko, V. N. (2024). Affinity Resins for the Isolation of Immunoglobulins G Obtained Using Biocatalytic Technology. International Journal of Molecular Sciences, 25(3), 1367. https://doi.org/10.3390/ijms25031367

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