基于细胞的噬菌体抗体库筛选技术  

Cell-based Screening of Phage Antibody Library

作　　者：尹长城[1] 黄华樑[2] 阎锡蕴[1] 

机构地区：[1]中国科学院生物物理研究所生物大分子国家重点实验室,北京100101 [2]北京安波特基因工程技术有限公司,北京102206

出　　处：《中国生物工程杂志》2008年第12期82-88,共7页China Biotechnology

基　　金：国家“863”计划(2006AA02A245)资助项目

摘　　要：肿瘤细胞表面抗原的表达量低,免疫原性弱,用固定化抗原方法筛选时,因抗原构象改变而很难获得有价值的抗体。同时,这类抗原多具有复杂的结构,其转运、定位与细胞类型及所处的微环境有关。在基于细胞的筛选中,靶抗原处于天然状态,且不需纯化,甚至可以对未知抗原进行筛选,因此广泛用于对内化抗体和血管内皮抗原表位抗体的筛选。基于细胞筛选的主要问题是非特异结合导致的选择背景过高,为此,许多研究都致力于改善选择的灵敏度和特异性。伴随高通量流式细胞术、组合化学及蛋白质组学研究技术的运用,细胞筛选技术将更趋实用和成熟。拟对细胞和组织的筛选技术作一概括,为了解基于细胞的筛选和优化实验设计提供参考。The limited number and low immunogenicity of tumor antigen make it difficult to screen out valuable phage antibody by panning on purified immobilized antigen because of the alternation of protein conformation. Most of these antigens have complex structure and the transporting and localization are depended on the cell types and microenvironment. Cell panning is applicable to complex antigen in its natural state without an additional purification procedure, or the exact antigen is not even known. So it is widely used in selection of tumor specific antibody, especially internalizing antibody and antibody specific targeting to vascular endothelial antigen. The major problem encountered in cell panning is related to the high background binding of non-specific antibody. Many efforts had been made to improve the specificity and sensitivity of cell panning. The combination of high throughout flow cytometry, combinatorial chemistry and proteome research will make this strategy more practical and fruitful. The progress of panning on intact cells and tissue sections was outlined to provide beneficial reference for understand and design of cell based panning.

关 键 词：抗体 细胞筛选 噬菌体展示 

分 类 号：Q819[生物学—生物工程]