An automated method for high-throughput protein purification applied to a comparison of His-tag and GST-tag affinity chromatography.

ABSTRACT:

Background

Functional Genomics, the systematic characterisation of the functions of an organism's genes, includes the study of the gene products, the proteins. Such studies require methods to express and purify these proteins in a parallel, time and cost effective manner.

Results

We developed a method for parallel expression and purification of recombinant proteins with a hexahistidine tag (His-tag) or glutathione S-transferase (GST)-tag from bacterial expression systems. Proteins are expressed in 96-well microplates and are purified by a fully automated procedure on a pipetting robot. Up to 90 microgram purified protein can be obtained from 1 ml microplate cultures. The procedure is readily reproducible and 96 proteins can be purified in approximately three hours. It avoids

SUBMITTER: Scheich C 

PROVIDER: S-EPMC183854 | biostudies-literature | 2003 Jul

REPOSITORIES: biostudies-literature