Preparation and functional analysis of the monoclonal antibodies against severe fever with thrombocytopenia syndrome bunyavirus structural proteins.
- Author:
Aqian LI
;
Lin LIU
;
Shuo ZHANG
;
Chuan LI
;
Quanfu ZHANG
;
Mifang LIANG
;
Dexin LI
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal;
immunology;
Antibodies, Viral;
immunology;
Antibody Specificity;
Bunyaviridae Infections;
immunology;
virology;
Female;
Humans;
Hybridomas;
immunology;
Mice;
Mice, Inbred BALB C;
Phlebovirus;
immunology;
Viral Structural Proteins;
immunology
- From:
Chinese Journal of Virology
2015;31(1):18-23
- CountryChina
- Language:Chinese
-
Abstract:
To prepare monoclonal antibodies (mAbs) against structural proteins of severe fever with thrombocytopenia syndrome bunyavirus (SFTSV), BALB/c mice were immunized using purified inactivated SFTSV virions as the antigens. Subsequently, hybridoma cell lines that secreted monoclonal antibodies against nucleoprotein (NP) and glycoproteins (GP) were obtained using a hybridoma technique. The antigen specificities of prepared mAbs were examined by indirect immunofluorescence and immunoprecipitation assays. Functional analyses were then performed,including the detection of IFA antibody titers,the levels of neutralizing activity and antibody affinities. After cell fusion and cloning,13 hybridoma cell lines secreted mAbs specifically against SFTSV-GP and 7 hybridoma cell lines secreted mAbs specifically against SFTSV-NP. Immunofluorescence and immunoprecipitation assays showed that the mAbs had high levels of antigen specificity. Among the 13 anti-SFTSV-GP mAbs,6 recognized Gn,whereas the others reacted with Gc. IFA titers of most anti-SFTSV-GP mAbs were between 1,280 and 20,480, and four anti-SFTSV-Gn mAbs showed neutralizing activity. Seven of the obtained anti-SFTSV-NP mAbs reacted specifically with NP,of which the IFA titers ranged from 5,120 to 20,480 with no observed neutralizing activity. Furthermore, two anti-SFTSV-GP mAbs, 1C8 and 1G8, showed high levels of affinity via a non-competitive ELISA. Our study lays the foundation for the development of further diagnostic assays and basic research into SFTSV.
