Cloning and production of human anti-B monoclonal antibody in ABO blood type

在我们的工作中，克隆了ABO血型人b抗原抗体可变区，并在大肠杆菌细胞中制备了重组Fab片段。Epstein-Barr病毒转化外周血淋巴细胞培养后，采用ELISA和流式细胞术筛选特异性抗体产生细胞。利用亚类特异性引物逆转录pcr构建cDNA文库，并在大肠杆菌表达系统中以质粒载体pFab-His2进行筛选。两个克隆在ELISA和血凝反应中与b抗原特异性反应。他们的抗b抗体重链和轻链核苷酸序列在免疫球蛋白亚群中分别为V_L1和V_H4。针对血型抗原结构的重组抗体可能具有表型适应性，这将为法医学应用提供有价值的手段。

In our work, cloning of the variable region of human antibodies directed to B-antigen in ABO blood group, and production of the recombinant Fab fragment in Escherichia coli cells were carried out. After the culture of peripheral lymphocyte transformed by Epstein-Barr virus, specific antibody-producing cells were screened by ELISA and flowcytometry. After reverse transcription-PCR using subclass-specific primers, cDNA library was constructed and screened with a plasmid vector, pFab-His2, in the E. coli expression system. Two clones specifically reacted with B-antigen in ELISA and hemagglutination. Their nucleotide sequence of the heavy and light chains of the anti-B human antibodies was determined to be V_L1 and V_H4, respectively, in the immunoglobulin subgroup. The recombinant antibodies to the blood group antigen structure may be adaptable for phenotyping, which will provide a valuable means for forensic applications.