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The neutralizing human recombinant antibodies to pathogenic Orthopoxviruses derived from a phage display immune library |
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| Authors: | Tikunova Nina Dubrovskaya Viktoriya Morozova Vera Yun Tatiana Khlusevich Yana Bormotov Nikolai Laman Aleksandr Brovko Fedor Shvalov Aleksandr Belanov Eugeni |
| Affiliation: | a Institute of Chemical Biology and Fundamental Medicine, Novosibirsk 630090, Russia b Federal State Research Center of Virology and Biotechnology “Vector”, Koltsovo, Novosibirsk reg. 630559, Russia c Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Pushchino, Moscow reg. 142290, Russia |
| Abstract: | A panel of recombinant human antibodies to orthopoxviruses was isolated from a combinatorial phage display library of human scFv antibodies constructed from the Vh and Vl genes cloned from the peripheral blood lymphocytes of Vaccinia virus (VACV) immune donors. Plaque-reduction neutralization tests showed that seven selected phage-displaying scFv antibodies (pdAbs) neutralized both CPXV and VACV, and five of them neutralized Monkeypox virus (MPXV). Western blot analysis of VACV and CPXV proteins demonstrated that seven neutralizing antibodies recognized a 35 kDa protein. To identify this target protein, we produced a recombinant J3L protein of CPXV and showed that all the selected neutralizing antibodies recognized this protein. Neutralizing pdAb b9 was converted into fully human mAb b9 (fh b9), and scFv b9 displayed high binding affinities (Kd of 0.7 and 3.2 nM). The fh b9 reduced VACV plaque formation in a dose-dependent manner. |
| Keywords: | Monkeypox virus Cowpox virus Vaccinia virus Neutralization Phage display ScFv Fully human mAb |
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