噬菌体
噬菌体展示
单克隆抗体
抗体
肽库
分子生物学
噬菌体
生物
抗体库
化学
病毒学
基因
生物化学
肽序列
遗传学
大肠杆菌
作者
David J. O’Connell,Baltazar Becerril,Arup Roy-Burman,Michael R. Daws,James D. Marks
标识
DOI:10.1016/s0022-2836(02)00561-2
摘要
Non-immune (naïve) phage antibody libraries have become an important source of antibodies for reagent, diagnostic, and therapeutic use. To date, reported naïve libraries have been constructed in phagemid vectors as fusions to pIII, yielding primarily single copy (monovalent) display of antibody fragments. For this work, we subcloned the single chain Fv (scFv) gene repertoire from a naïve phagemid antibody library into a true phage vector to create a multivalently displayed scFv phage library. Compared to monovalently displayed scFv, multivalent phage display resulted in improved efficiency of display as well as antibody selection. A greater number of antibodies were obtained and at earlier rounds of selection. Such increased efficiency allows the screening for binding antibodies after a single round of selection, greatly facilitating automation. Expression levels of antigen-binding scFv were also higher than from the phagemid library. In contrast, the affinities of scFv from the phage library were lower than from the phagemid library. This could be overcome by utilizing the scFv in a multivalent format, by affinity maturation, or by converting the library to monovalent display after the first round of selection.
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