从噬菌体抗体文库中筛选出的针对57-76 p21Ras中和表位的单链可变片段可识别亲本蛋白。

Single-chain variable fragments selected on the 57-76 p21Ras neutralising epitope from phage antibody libraries recognise the parental protein.

作者信息

Persic L, Horn I R, Rybak S, Cattaneo A, Hoogenboom H R, Bradbury A

机构信息

Laboratory of Molecular Neurobiology, International School of Advanced Studies (SISSA), Trieste, Italy.

出版信息

FEBS Lett. 1999 Jan 25;443(2):112-6. doi: 10.1016/s0014-5793(98)01684-6.

DOI:10.1016/s0014-5793(98)01684-6

PMID:9989586

Abstract

Phage antibodies have been widely prospected as an alternative to the use of monoclonal antibodies prepared by traditional means. Many monoclonal antibodies prepared against peptides are able to recognise the native proteins from which they were derived. Here we show that the same is also true for phage antibodies. We have selected a number of single-chain variable fragments (scFv) from a large phage scFv library against a peptide from the switch region II of p21Ras. This peptide is known to reside in a mobile area of the native protein and is the epitope of a well characterised monoclonal antibody. Selected scFvs were able to recognise native p21Ras in both ELISA and Western blots, indicating that peptides are also likely to be very useful in selecting from phage antibody libraries.

摘要

噬菌体抗体作为传统方法制备的单克隆抗体的替代物已得到广泛应用。许多针对肽段制备的单克隆抗体能够识别其来源的天然蛋白质。在此我们表明噬菌体抗体也是如此。我们从一个大型噬菌体单链可变片段（scFv）文库中筛选出了一些针对p21Ras开关区域II的肽段的scFv。已知该肽段位于天然蛋白质的一个可移动区域，并且是一种特性明确的单克隆抗体的表位。筛选出的scFv在酶联免疫吸附测定（ELISA）和蛋白质免疫印迹法（Western blot）中均能识别天然p21Ras，这表明肽段在从噬菌体抗体文库中进行筛选时可能也非常有用。