全人源抗狂犬病病毒单克隆抗体的筛选及功能验证

Screening and functional verification of human monoclonal antibodies against rabies virus

目的在噬菌体抗体颗粒水平及全分子抗体水平,对候选抗狂犬病病毒抗体进行筛选与抗体功能验证,以获得理想的备选抗体。方法通过固相包被法从全人源抗狂犬病病毒scFv噬菌体抗体库中筛选特异性抗体并对获得的抗体进行中和活性验证,进而挑选中和活性较高的抗体构建真核表达质粒,瞬时转染HEK293E细胞,对全分子抗体进行功能验证。结果对42株已获得的抗狂犬病病毒抗体在噬菌体抗体颗粒水平进行了中和活性验证,其中21株抗体RFFIT值>0.5 IU/mL;筛选出的4株抗体在全分子抗体水平进行中和活性验证,4株抗体RFFIT值>0.5 IU/mL。结论通过在噬菌体抗体颗粒水平及全分子抗体水平对候选抗狂犬病病毒抗体进行筛选与抗体功能验证,获得了4株具有明确中和活性的抗狂犬病病毒单克隆抗体。

Objective In order to obtain ideal candidate anti-rabies virus antibodies, candidate anti-rabies virus antibodies were screened and verified their antibody functions at phage antibody particles level and full molecular level. Methods The specific antibodies were screened from the whole human anti-rabies virus scFv phage antibody library by a solid-phase coating method, and the neutralizing activity of the obtained antibody was verified. Then antibodies with higher neutralizing activity were selected to construct eukaryotic expression plasmid, which was transiently transfected into HEK293 E cells to verify the function of the whole molecular antibody. Results The neutralizing activity of 42 obtained anti-rabies virus antibodies at phage antibody particle level was verified. Among them, the RFFIT value of 21 antibodies > 0.5 IU/mL;the neutralization activity of the screened four antibodies was verified at the whole molecular antibody level, the RFFIT value of the 4 antibodies > 0.5 IU/mL. Conclusion The candidate anti-rabies virus antibodies were screened and verified the antibody function at both phage antibody particles level and full molecular level, and four ideal monoclonal antibodies against rabies virus with defined neutralizing activity were obtained.