OBTAINING CHIMERIC VARIANTS HBcAg EXPOSING HIV-1 MPER FRAGMENTS
Andrey Pavlovich RUDOMETOV, Nadezhda Borisovna RUDOMETOVA, Boris Nikolaevich ZAYTSEV, Leonid Rudol’fovich LEBEDEV, Alexander Alekseevich ILYICHEV, Larisa Ivanovna KARPENKO
State Research Center of Virology and Biotechnology В«Vector» of Rospotrebnadzor
Keywords: ВИЧ-1, антигены, рекомбинантный НВсАg, рекомбинантные иммуногены, bNAbs, MPER, HIV-1, antigens, recombinant HBcAg, recombinant immunogens, bNAbs, MPER
Abstract
The HIV-1 epidemic is one
of the most acute global health problems. For several reasons, an
effective vaccine against this infection has not yet been created.
Currently, an important direction in the development of a vaccine
against HIV / AIDS is the design of immunogens that would be able to
induce antibodies that neutralize a high diversity of HIV-1 strains
(bNAbs). One approach to creating such immunogens is the construction of
chimeric virus-like particles (VLPs) exposing epitopes recognized by
bNAbs. The aim of the study was to obtain and characterize chimeric VLPs
based on HBcAg, exposing epitopes recognized by bNAbs 2F5 and 4E10.
Material and methods. The producing strains of chimeric HBcAg variants
were obtained by transforming E. coli BL21 cells with recombinant
plasmids carrying the HBcAg genes and containing insertions encoding
bNAbs epitopes 2F5 and 4E10. Purification of recombinant proteins was
performed using gel filtration on a sepharose CL-6B column. The ability
of recombinant HBcAg to form virus-like particles was assessed using
electron microscopy. Analysis of the antigenic properties of epitopes in
the composition of chimeric variants of HBcAg was performed using
immunoblotting. Results. A modified nucleotide sequence of the HBcAg
gene was obtained, which included the introduction of unique restriction
sites flanking the region of the main antigenic determinant of the
core. Based on this genetic construct, three recombinant plasmids
encoding chimeric HBcAg variants, including epitopes of bNAbs 2F5 and
4E10, were obtained. Using immunoblotting, it was found that epitopes
recognized by bNAbs retain their antigenic properties after insertion
into the HBcAg.
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