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使用活柱筛选特定噬菌体抗体。

Use of living columns to select specific phage antibodies.

作者信息

Bradbury A, Persic L, Werge T, Cattaneo A

机构信息

SISSA, Biophysics Sector, Trieste, Italy.

出版信息

Biotechnology (N Y). 1993 Dec;11(13):1565-9. doi: 10.1038/nbt1293-1565.

Abstract

Here we demonstrate that it is possible to confront two recombinant microorganisms in order to select one using the other. We have shown that an epitope derived from p21ras expressed within the outer membrane protein, LamB, can be recognized both by the monoclonal antibody Y13-259, as well as the single chain Fv fragment derived from it. This specificity, which is maintained when the Y13-259 single chain Fv is expressed as a fusion protein with the phage fd gene 3 protein, has allowed us to use the living column of LamB-ras to purify Y13-259 phage from a background of non-binding phage, even at dilutions as high as 10 phage in 10(10) irrelevant phage.

摘要

在此我们证明,为了选择一种重组微生物而使用另一种重组微生物是可行的。我们已经表明,在外膜蛋白LamB中表达的源自p21ras的表位,既能被单克隆抗体Y13 - 259识别,也能被源自它的单链Fv片段识别。当Y13 - 259单链Fv与噬菌体fd基因3蛋白作为融合蛋白表达时,这种特异性得以保持,这使我们能够利用LamB - ras的活柱从非结合噬菌体背景中纯化Y13 - 259噬菌体,即使在10¹⁰个无关噬菌体中有10个噬菌体的稀释度下也是如此。

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