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为了降低鼠源抗人CD3单抗的免疫原性,增加其在人体内的生物活性及治疗作用,使该抗体能更广泛更有效地长期多次用于人体治疗肿瘤、器官移植排斥反应及自身免疫性疾病.本文采用PCR技术从分泌抗CD3单抗的杂交瘤细胞HIT3a的mRNA中分离克隆了抗体的轻重链可变区基因的cDNA.以此轻重链可变区cDNA为特异探针从HIT3a基因文库中分离带有调控序列的功能性轻重链可变区基因;并将其插入到含有人κ轻链及人γ1重链恒定区基因的哺乳动物表达载体中成功地构建了抗人CD3人/鼠轻重链嵌合抗体基因,为研制人抗CD3人/鼠嵌合抗体完成了关键性的第一步.
In order to reduce the immunogenicity of murine anti-human CD3 monoclonal antibody and increase its bioactivity and therapeutic effect in human body, the antibody can be widely and more effectively used for treating human tumor, organ transplant rejection and self more widely Immune disease.In this paper, the cDNA of the light chain variable region (VHL) gene was isolated from the mRNA of HIT3a secreting anti-CD3 monoclonal antibody by PCR.This light chain variable region cDNA was used as a specific probe from HIT3a The functional light chain variable region genes with regulatory sequences were isolated from the gene library and inserted into the mammalian expression vector containing human kappa light chain and human γ1 heavy chain constant region gene to construct the anti-human CD3 / Mouse light chain heavy chain chimeric antibody gene for the development of human anti-CD3 human / mouse chimeric antibody to complete a crucial first step.