Obtaining and Characterization of the Monoclonal Antibodies Against G-Protein of the Respiratory Syncytial Virus

Abstract

The aim of this study was to obtain hybridomas producing monoclonal antibodies (Mabs) to the G-protein of the respiratory syncytial virus (RSV), and to evaluate their immunological characteristics and virus-neutralizing activity.Material and methods. Mouse Mabs were obtained using hybridoma technology. The properties of Mabs were studied by enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining (IF) of infected cells, as well as by biological neutralization test in vitro (NT). To identify epitopes recognized by the Mabs on G protein ELISA additivity test was used.Results. Hybridization of splenocytes with Sp2/0 myeloma cells and primary screening showed that 75 hybridomas produce antibodies interacting with purified virus, 17 of them also react with the recombinant G-protein in ELISA. In NT 4, hybridomas suppressed in vitro RSV infection by more than 50%. Cloning of these hybridomas revealed 4 monoclones producing the most active Mabs. Mab 1C11 was IgG2a, 3 others (5D4, 5G11 and 6H4) were IgM. Three IgM Mabs actively reacted with both RSV A2 and Long, and with G-protein; Mab 1C11 was less reactive with all antigens tested. All Mabs suppressed RSV infection, while Mab 5D4 supressed it almost completely (98%). IF analysis showed that all Mabs detected RSV G-protein in the cell cytoplasm, the largest number of infected cells was detected using Mab 5D4 (80%). It was shown that the isolated Mabs were directed to two non-overlapping epitopes on the RSV G-protein.Conclusion. The isolated Mabs can be used to detect RSV in clinical samples by ELISA and IF. The isolated Mabs can be used for humanized recombinant antibodies construction and for the treatment of RSV infection in future.

Antibodies as Biosensors’ Key Components: State-of-the-Art in Russia 2020–2021

“…Despite the general principle of synthesis and homology, the enzymes AlpA and AlpB differ in how they are secreted into the environment [51]. The mAbs described in [52] bind to two different epitopes of G protein of the respiratory syncytial virus, possess a virus-neutralizing activity, demonstrate an additive effect in interaction, detect the viral antigen in infected cell cultures, and can be used to reveal the virus in clinical material by immunofluorescence assay and ELISA. Studies using mAbs have shown the significance of the C-terminal domain of hemolysin II for the hemolytic activity of Bacillus cereus (which causes infections in persons with weakened immunity and is one of the widespread causes of intra-hospital infections) [53], as well as its ability to bind to cell membranes [54] and be oligomerized in the presence of cell membranes [55].…”

Obtaining and Characterization of the Monoclonal Antibodies Against G-Protein of the Respiratory Syncytial Virus

“…Эффективность паливизумаба у недоношенных детей с бронхолегочной дисплазией была подтверждена отечественными авторами в результате многоцентрового исследо-вания: у детей, которые относились к группе высокого риска тяжелого течения РСВ-инфекции по причине недоношенности, зарегистрировано снижение частоты госпитализаций после 3 инъекций паливизумаба до 0,3% [4]. Однако применение препарата не предотвращает повторного заболевания, эффективность препарата значительно ниже при лечении доношенных новорожденных и более старших детей, а более аффинные антитела к F-белку (мотавизумаб) вызывали аллергические реакции [5]. Кроме того, высокая стоимость препарата препятствует его широкому использованию.…”