Rapid detection of 6×-histidine-labeled recombinant proteins by immunochromatography using dye-labeled cellulose nanobeads

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• 作者：Eun-Sook Choi ; Se Geun Lee ; Sung-Jun Lee ; Eunjoo Kim
• 关键词：Dye ; labeled cellulose nanobeads ; Hexa ; histidine tag ; Immunochromatography ; Rapid test ; Recombinant protein detection
• 刊名：Biotechnology Letters
• 出版年：2015
• 出版时间：March 2015
• 年：2015
• 卷：37
• 期：3
• 页码：627-632
• 全文大小：660 KB
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• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Applied Microbiology
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-6776

文摘

A rapid and easy immunochromatography assay using dye-labeled cellulose nanobeads (CNBs) was developed to detect proteins with hexa-histidine tag (His-tag) to characterize recombinant proteins during purification. Recombinant ATG8 protein was used as a His-tagged protein, and ATG8-conjugated CNBs (A-CNBs) were prepared. The original ATG8 in the sample solution competed with A-CNBs for anti-His-tag antibodies spotted on to the strip resulting in an inverse relationship between ATG8 concentration and the colorimetric signal. The usefulness of this method was shown by adding ATG8 to a 1?% Escherichia coli extract. In addition, this assay can be used to detect other His-tagged proteins without protein-specific antibodies. Because the identification of fractions containing His-tagged proteins by western blotting or ELISA is labor-intensive and expensive, our method provides an efficient and cheaper alternative.