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以健康人的外周血淋巴细胞为来源 ,以偶联BSA的乙型肝炎病毒PreS1肽体外免疫 .分别从免疫和未经免疫的淋巴细胞提取RNA ,扩增抗体基因 ,构建大容量天然单链抗体 (scFv)噬菌体展示文库和体外免疫scFv抗体库 .以PreS1肽进行 3轮淘选后 ,抗原抗体反应结果显示 ,从免疫库中获得了亲和力 10 -7～ 10 -8M的抗乙型肝炎病毒PreS1的单链抗体 ,高于天然库的结果 (10 -6～ 10 -7M ) .测序结果表明两株抗体均为人抗体 .为基因工程抗体用于临床治疗乙型肝炎奠定基础 .同时证明淋巴细胞体外免疫方法构建的免疫抗体库优于大容量天然抗体库 .
In vitro, we used the Hepatitis B virus PreS1 peptide coupled with BSA to immunize in vitro from peripheral blood lymphocytes of healthy individuals.RNA was extracted from the immunized and unimmunized lymphocytes respectively to amplify the antibody gene and construct a large-capacity natural single-chain antibody (scFv) phage display library and in vitro immunized scFv antibody library.After three rounds of panning with PreS1 peptide, the results of antigen-antibody reaction showed that the anti-HBV PreS1 with affinity of 10 -7 -10 -8 M was obtained from the immune library (10 -6 ~ 10 -7 M) .The results of sequencing showed that both of the two antibodies were human antibodies, which laid the foundation for the clinical application of genetically engineered antibodies in the treatment of hepatitis B. At the same time, it was proved that lymphocytes in vitro The immune antibody library constructed by immune method is superior to the large-capacity natural antibody library.