血源性人免疫球蛋白-NK细胞重定向分子偶联物的构建与效应检测OA

Objective Based on the chemical coupling method,the blood-derived human immunoglobulin-NK cell retargeting molecular conjugate was constructed and its antiviral effect was evaluated.Methods Using human immunoglobulin products with high anti-VZV antibody titers as raw materials,the conjugates of human immunoglobulin and NK cell redirection molecule(anti-CD16a scFv)with different coupling degrees,IgG×αCD16a,were prepared by Traut's reagent and Sulfo-SMCC.The conjugated products were characterized by non-reducing SDS-PAGE,and the binding ability of IgG×αCD16a to effector cells(NK cells)was determined by flow cytometry.ADCC activity was detected by LDH release assay,the release of cell killing mediators(granzyme B,perforin)and cytokines(TNF-α,IFN-γ)was detected by ELISA.The killing ability of coupled product IgG×αCD16a and unconjugated human immunoglobulin on VZV infected cells was systematically evaluated.Results The coupling rate of IgG×αCD16a can reach more than 30％.IgG×αCD16a can bind to NK cells,and the binding ratio is higher than that of unconjugated human immunoglobulin.Compared with unconjugated human immunoglobulin,IgG×αCD16a significantly enhanced the ADCC activity on VZV infected cells and the release level of cytokines (TNF-α,IFN-γ). Conclusion The molecular chemical conjugate of blood-derived human immunoglobulin-NK cell redirection based on chemical coupling can effectively improve the antiviral effect of blood-derived human immunoglobulin,and provide a new idea for the development of a new generation of highly effective anti-viral antibody drugs.