Antibody arrays for high-throughput screening of antibody-antigen interactions

被引:510
|
作者
de Wildt, RMT
Mundy, CR
Gorick, BD
Tomlinson, IM
机构
[1] MRC, Mol Biol Lab, Cambridge CB2 2QH, England
[2] MRC, Ctr Prot Engn, Cambridge CB2 2QH, England
[3] UK Human Genome Mapping Project Resource Ctr, Cambridge CB10 1SB, England
关键词
antibody; array screening; high throughput; single-chain variable fragment (scFv); phage display;
D O I
10.1038/79494
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We have developed a novel technique for high-throughput screening of recombinant antibodies, based on the creation of antibody arrays. Our method uses robotic picking and high-density gridding of bacteria containing antibody genes followed by filter-based enzyme-linked immunosorbent assay (ELISA) screening to identify clones that express binding antibody fragments. By eliminating the need for liquid handling, we can thereby screen up to 18,342 different antibody clones at a time and, because the clones are arrayed from master stocks, the same antibodies can be double spotted and screened simultaneously against 15 different antigens. We have used our technique in several different applications, including isolating antibodies against impure proteins and complex antigens, where several rounds of phage display often fail. Our results indicate that antibody arrays can be used to identify differentially expressed proteins.
引用
收藏
页码:989 / 994
页数:6
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