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Generation and Characterization of Human Interferon-beta Neutralizing Humanized Antibody

Abstract

Humanization of antibodies for the development of novel therapeutic agents with low immunogenicity remains a topical problem in modern science. In the present work we describe the humanization of murine antibody B16 which binds and neutralizes human interferon-beta using the CDR-grafting method. Based on amino acid sequences of humanized and murine antibodies we constructed models of variable domains, analyzed, and compared them. The genes of humanized antibody hB16 and chimeric antibody chB16 were expressed in transient CHO cells. Antibodies were recovered from conditioned media, purified using affinity chromatography, and their properties were studied by biochemical and immunochemical methods. It was proven that humanized antibody hB16 possesses the same properties as murine mAb B16. This humanized antibody hB16 will be used in further work in order to obtain therapeutic immune complex composed of human interferon-beta and bispecific antibody which binds interferon-beta and the ErbB2 receptor.

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ACKNOWLEDGMENTS

The work was performed using the equipment of the Center for the collective use of ultrahigh-performance computing resources of Moscow State University.

Funding

The work was supported by the Ministry of Healthcare of the Russian Federation (agreement no. 075-15-2019-1385 of 19.06.2019, unique project identifier RFMEFI60417X0189).

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Correspondence to V. S. Rybchenko.

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Mononuclear peripheral blood cells were obtained from healthy donors’ blood. All donors provided voluntary informed consent.

Conflict of Interests

Authors declare they have no conflicts of interest.

Additional information

Translated by N. Onishchenko

Abbreviations: CDR, complementarity determining region; SDR, specificity determining region; FR, framework regions; VH and VL, variable domains of heavy and light chains of immunoglobulins; CH, constant domains of heavy chain of immunoglobulins; Cκ and Сλ, constant domains of light chains of immunoglobulins; mAbs, monoclonal antibodies; PBMC, peripheral blood mononuclear cells; IFN-β, interferon-beta; PBS(T), phosphate-buffered saline supplemented with Tween-20; MTT, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide; V, volume.

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Rybchenko, V.S., Panina, A.A., Novoseletsky, N.V. et al. Generation and Characterization of Human Interferon-beta Neutralizing Humanized Antibody. Russ J Bioorg Chem 46, 778–786 (2020). https://doi.org/10.1134/S1068162020050209

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  • DOI: https://doi.org/10.1134/S1068162020050209

Keywords:

  • humanized antibodies
  • chimeric antibodies
  • antibody structure modeling
  • СНО
  • interferon-beta