Abstract
We studied the possibility of using a broadly neutralizing anti-influenza A antibody as a module for the development of different protein constructs for diagnostics. For this purpose, we constructed two recombinant proteins—a Fab-fragment of the antibody and Fab-mCherry, which is a hybrid of the Fab-fragment and the mCherry fluorescent protein. Both proteins were expressed in Escherichia coli cells and purified in a functionally active state from culture medium. The antibody Fab-fragment was shown to bind all 11 tested strains of the influenza A H1N1 and H3N2 subtypes. A stronger binding was observed for group I hemagglutinins; this correlates with the immunochemical profile of the parental antibody. Comparison of the dissociation constants of complexes of the antibody Fab-fragment and Fab-mCherry with A(H1N1)/Solomon Islands/03/06 virus particles demonstrated that the attachment of the mCherry protein did not interfere with the antigen-binding properties of the antibody Fab-fragment.
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References
Graham-Rowe, D., Epidemiology: racing against the flu, Nature, 2011, vol. 480, no. 7376, pp. S2–S3.
Wrammert J., Koutsonanos D., Li G.M. et al. Broadly cross-reactive antibodies dominate the human B cell response against 2009 pandemic H1N1 influenza virus infection, J. Exp. Med., 2011, vol. 208, no. 1, pp. 181–193. Erratum in: J. Exp. Med., 2011, vol. 208, no. 2, p. 411.
Corti, D., Voss, J., Gamblin, S.J., et al., A neutralizing antibody selected from plasma cells that binds to group 1 and group 2 influenza A hemagglutinins, Science, 2011, vol. 333, no. 6044, pp. 850–856.
Deyev, S.M. and Lebedenko, E.N., Modern technologies for creating synthetic antibodies for clinical application, Acta Nat., 2009, vol. 1, no. 1, pp. 32–50.
Shaner, N.C., Campbell, R.E., Steinbach, P.A., Giepmans, B.N., Palmer, A.E., and Tsien, R.Y., Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein, Nat. Biotechnol., 2004, vol. 22, no. 12, pp. 1567–1572.
Kalinichenko, A.A., Toporova, V.A., Panina, A.A., Aliev, T.K., Kryukova, E.A., Shemchukova, O.B., Solopova, O.N., Pozdnyakova, L.P., Sveshnikov, P.G., Dolgikh, D.A., and Kirpichnikov, M.P., Development and characterization of antibodies against aflatoxins, Russ. J. Bioorg. Chem., 2010, vol. 36, no. 1, pp. 122–132.
Larina, M.V., Aliev, T.K., Solopova, O.N., Pozdnyakova, L.P., Korobova, S.V., Yakimov, S.A., Sveshnikov, P.G., Dolgikh, D.A., and Kirpichnikov, M.P., Neutralizing monoclonal and chimeric antibodies to human interferon, Russ. J. Bioorg. Chem., 2015, vol. 41, no. 3, pp. 316–326.
Studier, F.W., Protein production by auto-induction in high-density shaking cultures, Protein Expression Purif., 2005, vol. 41, no. 1, pp. 207–234.
Friguet, B., Chaffotte, A.F., Djavadi-Ohaniance, L., and Goldberg, M.E., Measurements of the true affinity constant in solution of antigen-antibody complexes by enzyme-linked immunosorbent assay, J. Immunol. Methods, 1985, vol. 77, no. 2, pp. 305–319.
Klotz, I.M., The Proteins, Klotz, I.M., Neurath, H., and Bailey, K., Eds., New York: Acad. Press, 1953, vol. 1.
Gross, L.A., Baird, G.S., Hoffman, R.C., Baldridge, K.K., and Tsien, R.Y., The structure of the chromophore whithin DsRed, a red fluorescent protein from coral, Proc. Natl. Acad. Sci. U. S. A., 2000, vol. 97, no. 22, pp. 11990–11995.
Feshchenko, E., Rhodes, D.G., Felberbaum, R., McPherson, C., Rininger, J.A., Post, P., and Cox, M.M.J., Pandemic influenza vaccine: characterization of A/California/07/2009 (H1N1) recombinant hemagglutinin protein and insights into H1N1 antigen stability, BMC Biotechnol., 2012, vol. 2012. doi 10.1186/ 1472-6750-12-77
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Original Russian Text © T.K. Aliev, I.G. Dement’yeva, V.A. Toporova, M.N. Bokov, L.P. Pozdnyakova, V.S. Rybchenko, D.A. Dolgikh, P.G. Sveshnikov, M.P. Kirpichnikov, 2016, published in Vestnik Moskovskogo Universiteta. Biologiya, 2016, No. 2, pp. 25–31.
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Aliev, T.K., Dement’yeva, I.G., Toporova, V.A. et al. Development and properties of recombinant proteins based on the broadly neutralizing antibody to influenza A virus. Moscow Univ. Biol.Sci. Bull. 71, 87–92 (2016). https://doi.org/10.3103/S0096392516020012
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DOI: https://doi.org/10.3103/S0096392516020012