The protein biotinylating agent, pentafluorophenyl 6-(biotinylamido)hexanoate (3) was synthesized. A straightforw ard synthetic route, biotin, biotin pentafluorophenyl ester (1), 6-(biotinylam ido)hexanoate (2) and pentafluorophenyl 6-(biotinylamido)hexanoate (3) was followed. This agent was used for biotin-labelling of different proteins, bovine serum albumin, immunoglobulin, horseradish peroxidase and β-galactosidase. Optimal conditions for the labelling were determined, as well as the influence of biotin-tagging to the biological activity of the proteins. An optimal ratio of the biotinylating agent to protein resulted in effective biotinylation without a significant decrease in the activity of the labelled enzyme. The labelled proteins proved to be suitable for immunodetection of proteins in both microplate and Western blot immunohistochemistry, as well as in a competitive steroid ELISA of testosterone and 20-hydroxyecdysone.
|Number of pages||6|
|Journal||Zeitschrift fur Naturforschung - Section B Journal of Chemical Sciences|
|Publication status||Published - Apr 1 1993|
- Biotin-Active Ester
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