全人源抗大别班达病毒抗体的制备及功能鉴定OA北大核心CSTPCD

Objective:To obtain Dabie bandavirus(DBV)specific antibody sequences from peripheral blood mononuclear cell(PBMC)of recovery patients with severe fever with thrombocytopenia syndrome(SFTS).Then fully human antibodies against DBV were prepared.Methods:Enzyme-linked immunosorbent assay(ELISA)was used to detect the plasma antibody of SFTS recovery patients.PBMC samples from the recovery patients were selected for magnetic bead enrichment,and the specific single B cells with DBV glycoprotein(DBV-Gn)protein binding activity were obtained by flow cytometry.The antibody sequences were obtained by B cell receptor sequencing,and the prokaryotic expression of single chain antibody was performed.Finally,the expression,binding capacity,and neutralizing activity of the antibodies were identified by Western blot,ELISA,and neutralization experiment in vitro.Results:Total 25 blood samples from SFTS recovery patients were collected.The antibodies showed binding activity with DBV-Gn and DBV nucleocapsid protein(DBV-NP)were detected in all plasma.Six types of single chain antibodies were obtained after flow cytometry sorting and BCR sequencing analysis,all of which had good binding capacity and neutralizing activity with DBV.Conclusion:Six fully human antibodies with neutralizing activity to DBV have been successfully expressed.