重组人抗HER2抗体在毕赤酵母中的表达及活性研究
发布时间：2018-07-26 12:19
【摘要】： 本研究利用基因重组技术，在已经构建出的具有Fc段的完全人源抗狂犬病毒抗体表达载体基础上，通过PCR介导的定点突变技术，构建出一个可供CDR3自由筛选、鉴定、表达的通用载体。 将体外合成的人源、鼠源抗HER2抗体的CDR3连接至构建出的通用载体上，通过对其在酵母中表达产物的生物学及免疫学功能的检测，对本实验所构建出的通用载体和表达出的重组人源、鼠源抗HER2抗体进行鉴定。 本研究还利用网络数据库，以首个应用于HER2阳性乳腺癌患者的人源化HER2重组单克隆抗体Herceptin作为参照，对重组人源、鼠源抗HER2抗体分别进行一级结构、二级结构和三维结构分析，通过三者之间的对比，找出决定HER2抗体活性的关键，为HER2治疗性单抗的研制提供理论和实验依据。
[Abstract]:In this study, by using gene recombination technique, based on the constructed full-human anti-rabies virus antibody expression vector with FC segment and PCR mediated site-directed mutagenesis, a CDR3 free screening and identification was constructed. A universal vector for expression. The CDR3 of human and mouse anti HER2 antibody synthesized in vitro was ligated to the constructed universal vector, and the biological and immunological functions of the expressed product in yeast were detected. The universal vector constructed in this experiment and the recombinant human and mouse anti-HER2 antibody were identified. Using the network database, the first humanized HER2 recombinant monoclonal antibody (Herceptin) applied to HER2 positive breast cancer patients was used as the reference, and the primary structure, secondary structure and three dimensional structure of the recombinant human and murine anti-HER2 antibodies were analyzed respectively. The key to determine the activity of HER2 antibody was found out through the comparison among them, which provided theoretical and experimental basis for the development of HER2 therapeutic monoclonal antibody.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392.11
本文编号：2145990
[Abstract]:In this study, by using gene recombination technique, based on the constructed full-human anti-rabies virus antibody expression vector with FC segment and PCR mediated site-directed mutagenesis, a CDR3 free screening and identification was constructed. A universal vector for expression. The CDR3 of human and mouse anti HER2 antibody synthesized in vitro was ligated to the constructed universal vector, and the biological and immunological functions of the expressed product in yeast were detected. The universal vector constructed in this experiment and the recombinant human and mouse anti-HER2 antibody were identified. Using the network database, the first humanized HER2 recombinant monoclonal antibody (Herceptin) applied to HER2 positive breast cancer patients was used as the reference, and the primary structure, secondary structure and three dimensional structure of the recombinant human and murine anti-HER2 antibodies were analyzed respectively. The key to determine the activity of HER2 antibody was found out through the comparison among them, which provided theoretical and experimental basis for the development of HER2 therapeutic monoclonal antibody.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392.11
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