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Vol 35(2019) N 2 p. 49-57; DOI 10.21519/0234-2758-2019-35-2-49-57
E.N. Kosobokova1*, E.V. Sheshukova2, M.V. Pinyugina1, K.M. Konoplina1, V.S. Kosorukov1

Development of Fusion Protein Produce Technology Based on a Humanized Monoclonal Antibody Specific to Tumor Antigen HER2 and a Recombinant Human Interferon-α-2b in Nicotiana benthamiana

1Blokhin National Medical Research Center of Oncology, 115478 Moscow, Russia
2Vavilov Institute of General Genetics of Rusian Academy Scinces, 119991 Moscow, Russia

*ekkos@mail.ru
Received - 21.10.2018; Accepted - 15.03.2019

References

Fusion proteins based on a recombinant human interferon-α-2b and a humanized monoclonal antibody specific to tumor antigen HER2 have been obtained by transient expression in Nicotiana benthamiana. To provide an effective expression and a proper immunocytokine molecule assembly the genetic constructs optimization was performed. The expression level was shown to depend on the linker length between the cytokine and the antibody moieties in the hybrid molecule. The modification of the interferon domain sequence by replacing cysteine residues with serine showed a slight increase in the production level of the target protein. The assembly of full-size fusion protein molecules was confirmed by the qualitative method of «combined ELISA» developed by the authors. The preservation of the affine domain function in the fusion proteins was proved by the ELISA method in interaction with the antigen.

interferon- α-2b, antigen HER2, plant transient expression, recombinant fusion proteins.



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