ANTIBODY AS A SURROGATE RECEPTOR IN THE SCREENING OF A PHAGE DISPLAY LIBRARY
ANTIBODY AS A SURROGATE RECEPTOR IN THE SCREENING OF A PHAGE DISPLAY LIBRARY
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DOI：
10.1016/0378-1119(93)90154-u
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发表时间：
1993-06-15
期刊：
影响因子：
3.5
通讯作者：
WINTER, J
中科院分区：
文献类型：
--
作者：
ROBERTS, D;GUEGLER, K;WINTER, J
抗生物素多克隆抗体 (Ab)（抗生物素抗体）被评估为链霉亲和素 (Sv) 生物素结合袋的形状模拟来源。用抗生物素淘选 6 聚体噬菌体展示文库，并将选定的肽序列与直接用 Sv 淘选获得的序列进行比较。使用抗生物素抗体探针鉴定共有 XXYYLH，而 GDWVFI 和 PWPWLG 是通过 Sv 淘选鉴定的主要基序。尽管在两次淘选的产物序列之间观察到一些相似性，但只有展示 GDWVFI 和 PWPWLG 肽的噬菌体在 ELISA 和微淘选测定中证明了生物素敏感的 Sv 结合。展示 GDWVFI 的噬菌体可以用酸或生物素从 Sv 中洗脱，但不能用磷酸盐缓冲盐水 (PBS) 洗脱，而展示 PWPWLG 的噬菌体用酸、生物素和 PBS 洗脱。
Polyclonal antibody (Ab) against biotin (anti-biotin Ab) was evaluated as a source of shape mimics for the biotin-binding pocket of streptavidin (Sv). A 6-mer phage display library was panned with anti-biotin and the selected peptide sequences compared to those obtained by panning directly with Sv. The consensus XXYYLH was identified with the anti-biotin Ab probe, while GDWVFI and PWPWLG were the major motifs identified by Sv panning. Although some similarities were observed between sequences of the products from both pannings, only phage displaying the GDWVFI and PWPWLG peptides demonstrated biotin-sensitive Sv binding in ELISA and micropanning assays. Phage displaying GDWVFI could be eluted from Sv with either acid or biotin, but not phosphate-buffered saline (PBS), whereas phage displaying PWPWLG eluted with acid, biotin and PBS.