噬菌体展示
化学
噬菌体
抗体
细胞生物学
生物化学
分子生物学
单克隆抗体
噬菌体
生物
淘选
重组DNA
计算生物学
肽库
作者
Isidro Hötzel,Vicki Chiang,Jingyu Diao,Homer Pantua,Henry R. Maun,Sharookh B. Kapadia
出处
期刊:Protein Engineering Design & Selection
[Oxford University Press]
日期:2011-09-01
卷期号:24 (9): 679-689
被引量:36
标识
DOI:10.1093/protein/gzr039
摘要
The application of phage display technology to mammalian proteins with multiple transmembrane regions has had limited success due to the difficulty in generating these proteins in sufficient amounts and purity. We report here a method that can be easily and generally applied to sorting of phage display libraries with multispan protein targets solubilized in detergent. A key feature of this approach is the production of biotinylated multispan proteins in virions of a baculovirus vector that allows library panning without prior purification of the target protein. We obtained Fab fragments from a naive synthetic antibody phage library that, when engineered into full-length immunoglobulin (Ig)G, specifically bind cells expressing claudin-1, a protein with four transmembrane regions that is used as an entry co-receptor by the hepatitis C virus (HCV). Affinity-matured variants of one of these antibodies efficiently inhibited HCV infection. The use of baculovirus particles as a source of mammalian multispan protein facilitates the application of phage display to this difficult class of proteins.
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