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Generation and Characterization of Human Interferon-beta Neutralizing Humanized Antibody

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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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REFERENCES

  1. Hwang, W.Y. and Foote, J., Methods, 2005, vol. 36, pp. 3–10. https://doi.org/10.1016/j.ymeth.2005.01.001

    Article  CAS  PubMed  Google Scholar 

  2. Scott, A.M., Lee, F., Hopkins, W., Cebon, J.S., Wheatley, J.M., and Liu, Z., J. Clin. Oncol., 2001, vol. 19, pp. 3976–3987. https://doi.org/10.1200/JCO.2001.19.19.3976

    Article  CAS  PubMed  Google Scholar 

  3. Buist, M.R., Kenemans, P., van Kamp, G.J., and Haisma, H.J., Cancer Immunol. Immunother., 1995, vol. 40, pp. 24–30. https://doi.org/10.1007/BF01517232

    Article  CAS  PubMed  Google Scholar 

  4. Roque-Navarro, L., Mateo, C., Lombardero, J., Mustelier, G., Fernández, A., Sosa, S., Morrison, S.L., and Pérez, R., Hybridoma Hybridomics, 2004, vol. 22, pp. 245–257. https://doi.org/10.1089/153685903322328974

    Article  CAS  Google Scholar 

  5. Richards, J., Auger, J., Peace, D., Gale, D., Michel, J., Koons, A., Haverty, T., Zivin, R., Jolliffe, L., and Bluestone, J.A., Cancer Res., 1999, vol. 59, pp. 2096–2101.

    CAS  PubMed  Google Scholar 

  6. Almagro, J. and Franson, J., BioScience, 2008, vol. 13, pp. 1619–1633.

    CAS  Google Scholar 

  7. Hale, G. and Phillips, M., Biochem. Soc. Transact., 1995, vol. 23, pp. 1057–1063. https://doi.org/10.1042/bst0231057

    Article  CAS  Google Scholar 

  8. Safdari, Y., Farajniaa, S., Asgharzadehb, M., and Khalili, M., Biotech. Genet. Eng. Rev., 2013, vol. 29, pp. 175–186. https://doi.org/10.1080/02648725.2013.801235

    Article  CAS  Google Scholar 

  9. Makabe, K., Nakanishi, N., Tsumoto, K., Tanaka, Y., Kondo, H., Umetsu, M., Sone, Y., Asano, R., and Kumagai, I., J. Biol. Chem., 2008, vol. 283, pp. 1156–1166. https://doi.org/10.1074/jbc.M706190200

    Article  CAS  PubMed  Google Scholar 

  10. Wedemayer, G.J., Patten, PA., Wang, L.H., Schultz, P.G., and Stevens, R.C., Science, 1997, vol. 276, pp. 1665–1669.

    Article  CAS  Google Scholar 

  11. Zimmermann, J., Oakman, E.L., Thorpe, I.F., Shi, X., Abbyad, P., Brooks, C.L., Boxer, S.G., and Romesberg, F.E., Proc. Natl. Acad. Sci. U. S. A., 2006, vol. 103, pp. 13 722–13 727.

    Article  Google Scholar 

  12. Kabat, E.A., Sequences of Immunological Interest, 5 ed., Bethesda, Md, USA: Public Health Service, NIH, 1991.

    Google Scholar 

  13. Ilina, E.N., Solopova, O.N., Balabashin, D.S., Larina, M.V., Aliev, T.K., Grebennikova, T.V., Losich, M.A., Zaykova, O.N., Sveshnikov, P.G., Dolgikh, D.A., and Kirpichnikov, M.P., Russ. J. Bioorg. Chem., 2019, vol. 45, pp. 59–68. https://doi.org/10.1134/S0132342319010081

    Article  Google Scholar 

  14. Rizner, T.L., Biochem. Mol. Biol. Educ., 2014, vol. 42, pp. 152–159. https://doi.org/10.1002/bmb.20764

    Article  CAS  PubMed  Google Scholar 

  15. Friguet, B., Chaffotte, A.F., Djavadi-Ohaniance, L., and Goldberg, M.E., J. Immunol. Methods, 1985, vol. 77, pp. 305–319.

    Article  CAS  Google Scholar 

  16. https://cdn.gelifesciences.com/dmm3bwsv3/AssetStream. aspx?mediaformatid=10061&destinationid=10016& assetid=14782

  17. Supino, R., In vitro Toxicity Testing Protocols, Humana Press, 1995, pp. 137–149.

    Google Scholar 

  18. Laemmli, U.K., Nature, 1970, vol. 227, pp. 680–685. https://doi.org/10.1038/227680a0

    Article  CAS  PubMed  Google Scholar 

  19. www.thermofisher.com/order/catalog/product/15596026#/ 15596026.

  20. Weitzner, B.D., Jeliazkov, J.R., Lyskov, S., Marze, N., Kuroda, D., Frick, R., Bryfogle, A.A., Biswas, N., and Gray, J.J., Nat. Protoc., 2017, vol. 12, pp. 401–416. https://doi.org/10.1038/nprot.2016.180

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Abraham, M.J., Murtola, T., Schulz, R., Pall, S., Smith, J.C., Hess, B., and Lindahl, E., SoftwareX, 2015, vols. 1–2, pp. 19–25. https://doi.org/10.1016/j.softx.2015.06.001

    Article  Google Scholar 

  22. Humphrey, W., Dalke, A., and Schulten, K., J. Mol. Graph., 1996, vol. 14, p. 3338.

    Article  Google Scholar 

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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).

Author information

Authors and Affiliations

  1. Faculty of Biology, Moscow State University, 119192, Moscow, Russia

    V. S. Rybchenko, N. V. Novoseletsky, D. A. Dolgikh & M. P. Kirpichnikov

  2. Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, 117997, Moscow, Russia

    A. A. Panina, D. S. Balabashin, S. A. Yakimov, D. A. Dolgikh & M. P. Kirpichnikov

  3. Russian Research Center for Molecular Diagnostics and Therapy, 117638, Moscow, Russia

    O. N. Solopova & P. G. Sveshnikov

  4. Chemistry Department, Moscow State University, 119234, Moscow, Russia

    T. K. Aliev

  5. Blokhin National Medical Research Center of Oncology, Ministry of Health of the Russian Federation, 115478, Moscow, Russia

    O. N. Solopova

Authors
  1. V. S. Rybchenko
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  2. A. A. Panina
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  3. N. V. Novoseletsky
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  4. D. S. Balabashin
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  5. S. A. Yakimov
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  6. O. N. Solopova
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  7. P. G. Sveshnikov
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  8. T. K. Aliev
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  9. D. A. Dolgikh
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  10. M. P. Kirpichnikov
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Corresponding author

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.

Corresponding author: phone: +7 (977) 272-87-63; e-mail: vl-adislavrusia@yandex.ru.

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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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