Integrate System for Creation, Detection and Production of Antibodies with Variety of Characteristics

在药物开发、医学诊断分析和生物过程、各种抗体的生产、抗体与蛋白质相互作用的检测、抗体的筛选方法和有效的抗体生产过程中是必不可少的。在本研究的基础上，我们发展了一种简便、快速的抗体配基的获得方法，用于识别和检测目的蛋白。1.利用抗原吸附PVDF膜和脂质体免疫印迹技术，建立了一种检测大肠杆菌分泌抗体的菌落提升法。利用HRP包被的免疫脂质体和疏水膜，建立了一种灵敏度高两个数量级的免疫印迹分析方法。3.通过H链和L链文库转化的酵母细胞交配，获得了大量的全抗体库，并建立了酵母细胞分泌产生抗体的方法。4.建立了一种利用免疫脂质体的亲和筛选方法，并将其应用于生物筛选。

In the fields of development of pharmaceuticals, medical diagnosis-analysis and bioprocesses, production of variety of antibodies, detection of the interactions between antibody and protein, screening methods of antibodies and effective production processes of antibodies are essential. In this work, we developed the simple and rapid methods to obtain antibody ligands for recognition and detection of target proteins, based on our recent researches on production of large libraries of antibodies, a high sensitivity detection method of proteins and effective production of antibodies by secretion from yeast cells.1.A colony lift assay for detection of antibody secreted from E. coli was developed by use of antigen-adsorbed PVDF membrane and liposome immunoblotting assay. By use of this method, high density and micro-assay was possible and the number of samples treated was increased.2.By use of HRP encapsulating immunoliposomes and hydrophobic membranes, an immunoblotting assay method with two orders of magnitude higher sensitivity, compared with the conventional immunoblotting assay using HRP-conjugated antibody, was developed.3.A large library of whole antibodies was obtained based on mating of yeast cells transformed by libraries of H- and L-chains, and aproduction method of antibodies by secretion from yeast cells was established.4.An affinity biopanning method utilizing immunoliposomes was developed and it was applied to biopanning selection with a high efficiency.