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Rapid detection of 6x-histidine-labeled recombinant proteins by immunochromatography using dye-labeled cellulose nanobeads
- Rapid detection of 6x-histidine-labeled recombinant proteins by immunochromatography using dye-labeled cellulose nanobeads
- Choi, Eun-Sook; Lee, Se Geun; Lee, Sung-Jun; Kim, Eunjoo
- DGIST Authors
- Lee, Se Geun; Lee, Sung-Jun; Kim, Eunjoo
- Issue Date
- Biotechnology Letters, 37(3), 627-632
- Article Type
- Amino Acids; Antibodies; Cellulose; Chemical Analysis; Chemistry Techniques, Analytical; Colorimetry; Dye-Labeled Cellulose Nanobeads; Escherichia Coli; Hexa-Histidine Tag; His-Tagged Proteins; Hybrid Protein; Immunoaffinity Chromatography; Immunochromatography; Immunochromatography Assays; Inverse Relationship; Metabolism; Nano-Structured Materials; Nanobeads; Nanoparticle; Nanoparticles; Procedures; Rapid Test; Recombinant Fusion Proteins; Recombinant Protein Detection; Recombinant Proteins; Time; Time Factors; Western Blotting
- 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. © 2014, Springer Science+Business Media Dordrecht.
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