論文

査読有り
2016年2月

Application of preparative disk gel electrophoresis for antigen purification from inclusion bodies

PROTEIN EXPRESSION AND PURIFICATION
  • Yuki Okegawa
  • ,
  • Masanori Koshino
  • ,
  • Teruya Okushima
  • ,
  • Ken Motohashi

118
開始ページ
77
終了ページ
82
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1016/j.pep.2015.10.008
出版者・発行元
ACADEMIC PRESS INC ELSEVIER SCIENCE

Specific antibodies are a reliable tool to examine protein expression patterns and to determine the protein localizations within cells. Generally, recombinant proteins are used as antigens for specific antibody production. However, recombinant proteins from mammals and plants are often overexpressed as insoluble inclusion bodies in Escherichia coli. Solubilization of these inclusion bodies is desirable because soluble antigens are more suitable for injection into animals to be immunized. Furthermore, highly purified proteins are also required for specific antibody production. Plastidic acetyl-CoA carboxylase (ACCase: EC 6.4.1.2) from Arabidopsis thaliana, which catalyzes the formation of malonyl-CoA from acetyl-CoA in chloroplasts, formed inclusion bodies when the recombinant protein was overexpressed in E. coli. To obtain the purified protein to use as an antigen, we applied preparative disk gel electrophoresis for protein purification from inclusion bodies. This method is suitable for antigen preparation from inclusion bodies because the purified protein is recovered as a soluble fraction in electrode running buffer containing 0.1% sodium dodecyl sulfate that can be directly injected into immune animals, and it can be used for large-scale antigen preparation (several tens of milligrams). (C) 2015 Elsevier Inc. All rights reserved.

Web of Science ® 被引用回数 : 3

リンク情報
DOI
https://doi.org/10.1016/j.pep.2015.10.008
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/26494602
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000367424600011&DestApp=WOS_CPL

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