High-throughput solubility assay for purified recombinant protein immunogens

被引：15

作者：

Stenvall, M ^{[1
]}

Steen, J ^{[1
]}

Uhlén, M ^{[1
]}

Hober, S ^{[1
]}

Ottosson, J ^{[1
]}

机构：

[1] Royal Inst Technol, AlbaNova Univ Ctr, Dept Biotechnol, S-10691 Stockholm, Sweden

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS | 2005年 / 1752卷 / 01期

关键词：

protein solubility; high-throughput; in vitro assay; urea;

D O I：

10.1016/j.bbapap.2005.07.002

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A high-throughput assay is described for analysis of the solubility of purified recombinant proteins. The assay is based on affinity purification of proteins in the presence of chaotropic agents followed by a dilution and incubation step to investigate the solubility in the absence of high concentrations of such agents. The assay can be performed in a 96-well format, which makes it well suited for high-throughput applications. For 125 recombinant proteins expressed as part of an antibody-based proteomics effort, experimental solubility data were compared to calculated hydrophobicity values based on the amino acid sequence of each protein. This comparison showed only weak correlation between the theoretical and experimental values, which emphasizes the importance of experimental assays to determine the solubility of recombinant proteins. (c) 2005 Elsevier B.V All rights reserved.

引用

页码：6 / 10

页数：5

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