2019新型冠状病毒治疗性中和抗体的快速研发和作用机制研究

SARS-CoV-2通过ACE2受体感染细胞，如何迅速有效地找到中和抗体，阻断细胞感染途径，是重要治疗手段。传统中和抗体的制备复杂耗时，需借助患者血清和感染膜区相关假病毒进行动物实验才能得到中和抗体产生细胞。我们前期建立了Beacon数字化单细胞光导平台获取乙肝患者中和抗体的方法。在该课题中我们沿用此方法从COVID-19患者外周血B淋巴细胞筛选出高特异性的SARS-CoV-2 S蛋白特异抗体B细胞，利用多重荧光技术对细胞进行功能验证，通过野生型和点突变ACE2的293T-ACE2细胞比较不同单克隆抗体对SARS-CoV-2假病毒感染的阻断作用，进而获取SARS-CoV-2抗体B细胞，进行克隆表达，最终得到高效SARS-CoV-2诊断和治疗抗体，并探讨ACE2对SARS-CoV-2中和作用影响机制，为救治生命提供重要保障。

SARS-CoV-2 virus infects cells through ACE2 receptor. How to quickly and effectively find neutralizing antibodies, block the cell infection pathway, is an important treatment. The preparation of traditional neutralizing antibody is complex and time-consuming, so it requires animal experiments and serial studies that are carried out with the help of patient serum and related pseudoviruses in the infected membrane area to obtain neutralizing antibody producing cells. In the early stage, we established Beacon digital single cell photoconductive platform to obtain neutralizing antibody of hepatitis B patients. In this project, we use this method to screen highly specific SARS-CoV-2 S protein specific antibody B cells from B lymphocytes of peripheral blood of patients with COVID-19, and use multiple fluorescence technology to verify the function of the cells.Through wild type and ACE2 point mutation with the 293T-ACE2 cells, we compare the blocking effect of different monoclone antibodies on SARS-CoV-2 pseudovirus infection and then obtain SARS-CoV-2 antibody B cells with clone expression, finally, acquire the highly effective SARS-CoV-2 diagnostic and therapeutic antibody.In addition, we will explore the influence mechanism of ACE2 on SARS-CoV-2 neutralization , so as to provide an important guarantee for the treatment of life.