Iterative size-exclusion chromatography coupled with liquid chromatographic mass spectrometry to enrich and identify tight-binding ligands from complex mixtures

被引：13

作者：

Davis, RG ^{[1
]}

Anderegg, RJ ^{[1
]}

Blanchard, SG ^{[1
]}

机构：

[1] Glaxo Wellcome Res Labs, Res Triangle Pk, NC 27709 USA

来源：

TETRAHEDRON | 1999年 / 55卷 / 39期

关键词：

chromatography; combinatorial chemistry; mass spectrometry; proteins;

D O I：

10.1016/S0040-4020(99)00685-7

中图分类号：

O62 [有机化学];

学科分类号：

070303 ; 081704 ;

摘要：

A method is described for the enrichment of tight-binding ligands from complex mixtures. The mixture is equilibrated with a protein or receptor, and protein:ligand complexes are separated from unbound ligands by size-exclusion. If the resulting mixture is allowed to re-establish its equilibrium and then passed again through the size-exclusion column, significant enrichment of tight-binding ligands is realized, even in cases where the weak-binders are present in large excess. Ligands are identified by liquid chromatography and mass spectrometry following the size-exclusion step. The process can be repeated multiple times to further enrich ligands. Analysis of pools of ligands generated by combinatorial chemistry is an obvious application of the technique. (C) 1999 Published by Elsevier Science Ltd. All rights reserved.

引用

页码：11653 / 11667

页数：15

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