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Generation of Hybridomas Producing Monoclonal Antibodies to the NS1 Protein of the Zika Virus

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Abstract

Hybridomas producing monoclonal antibodies (mABs) to the recombinant protein NS1 of the Zika virus synthesized in human embryonic kidney HEK293 (ZV293) cells have been obtained. The specificity of antibodies has been examined by the enzyme-linked immunosorbent assay (ELISA) using the similarly synthesized NS1 proteins of flaviviruses: the Dengue 1–4 serotype virus, Japanese encephalitis virus, yellow fever virus, tick-borne encephalitis virus, and the West Nile fever virus. Two types of hybridomas whose mABs interact with thermolabile (mABs 2F6 and 3F3) and thermostable epitopes (mABs 1B9, 6D7, and 6F2) of the NS1ZV protein have been selected. It has been shown that the 3F3 mAB, which is specific to the thermolabile determinant, can be used as both binding and detecting antibodies in the sandwich ELISA. Based on the 3F3 mAB, an experimental immunochromatographic test for the detection of the NS1ZV antigen has been developed. It has also been found that mABs 1B9, 6D7, and 6F2 against thermostable epitopes of the NS1 protein specifically recognize the NS1ZV protein after its heating for 15 min to 98°C.

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Abbreviations

BSA:

bovine serum albumin

ELISA:

enzymelinked immunosorbent assay

IC test:

immunochromatographic test

CG:

colloid gold

mAB:

monoclonal antibody

RT-PCR:

reverse transcription polymerase chain reaction

PEG:

polyethylene glycol

PBS–T:

0.1 M phosphate-buffered saline with 0.5% Twin-20

HAT:

a mixture of hypoxanthine, aminopterin, and thymidine

НТ:

a mixture of hypoxanthine and thymidine

NR:

neutralization reaction: NS1ZV293, the recombinant protein NS1 of the Zika virus, synthesized in human embryonic kidney cell line HEK293

NS1ZVE. сoli :

the recombinant protein NS1 of the Zika virus, synthesized in E. сoli

ZVVero:

lysate of Vero cells infected with the Zika virus

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Vetchinin, S.S., Shevyakov, A.G., Fedyukina, G.N. et al. Generation of Hybridomas Producing Monoclonal Antibodies to the NS1 Protein of the Zika Virus. Russ J Bioorg Chem 44, 745–754 (2018). https://doi.org/10.1134/S1068162019010187

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  • DOI: https://doi.org/10.1134/S1068162019010187

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