PMID: 3772022Sep 1, 1986Paper

An improved method for the single-step purification of streptavidin

Journal of Biochemical and Biophysical Methods
E A BayerM Wilchek

Abstract

A new method for the preparation of a more efficient, stable iminobiotin-containing resin for the isolation of streptavidin was developed. CL-Sepharose was activated with p-nitrophenyl chloroformate, and the resultant carbonate derivative was reacted with diaminohexane. Subsequent reaction of the amino-containing resin with iminobiotin-N-hydroxysuccinimide ester (in an organic solvent) yielded the stable affinity resin. The capacity of this resin for either avidin or streptavidin was 12 mg per ml resin, and streptavidin could be purified in one step directly from the culture broth of Streptomyces avidinii. The biotin-binding protein isolated in this manner exhibited a major band at about 75 kDa and a minor band at about 150 kDa. Under denaturing conditions, a spectrum of subunit molecular weights ranging between 15 and 19 kDa was detected, the distribution of which depended upon the specific preparation.

References

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Citations

Dec 1, 1990·Journal of Protein Chemistry·G P KurzbanP M Horowitz
Jun 1, 1990·Journal of Industrial Microbiology·L AldwinJ Hunter
Oct 4, 1988·Journal of Immunological Methods·J CazinJ E Butler
Nov 1, 1992·Journal of Chemical Neuroanatomy·G BrücknerD Biesold
May 27, 1999·Journal of Biochemical and Biophysical Methods·Z Gan, R R Marquardt
Jan 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·T Sano, C R Cantor
Mar 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·T SanoC R Cantor
Sep 25, 1999·Protein Expression and Purification·K J AirenneM S Kulomaa
Aug 16, 1990·Biochemical and Biophysical Research Communications·R AlonM Wilchek
Nov 17, 2015·Chemistry : a European Journal·Elise VilleminEric Doris
Jan 1, 1990·Biomaterials, Artificial Cells, and Artificial Organs·C RupprichA Könnecke
Nov 9, 1990·Journal of Chromatography·W D SchwidopR Claus
Mar 6, 1999·Journal of Chromatography. B, Biomedical Sciences and Applications·J Turková
Oct 1, 1989·Trends in Biochemical Sciences·M Wilchek, E A Bayer

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