Methods for a quantitative release of biotinylated peptides and proteins from streptavidin complexes
Biotinylated molecules, like proteins or peptides, DNA, RNA and PNA are quantitatively released from streptavidin complexes by adding a mixture of SX. After elution and removal of SX, e.g. by evaporation, the biotinylated molecules can be used for subsequent applications and analyses.
Due to the extraordinary stability of the biotinstreptavidin complex, biotin labeling of organic and inorganic molecules has emerged to a commonly used technique for numerous applications. The interaction is unaffected by wide extremes of pH, temperature, organic solvents and other denaturing agents.
The extraordinary stability of the biotin-streptavidin complex poses a major problem when biotinylated molecules need to be recovered from such complexes. Harsh elution conditions are required to dissociate the complex using heat and chemical reagents like SDS which are not compatible with subsequent analytical techniques especially mass spectrometry.
To circumvent these restrictions DKFZ inventors have developed a simple and mild method for complete dissociation of biotin-streptavidin complexes.
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The biotinylated molecules, like proteins or peptides, DNA, RNA and PNA are quantitatively released from streptavidin complexes by simply adding a mixture of SX (a well-known, low priced, save, and easy to handle liquid substance). After elution and removal of SX, e.g. by evaporation, the biotinylated molecules can be used for subsequent applications and analyses.
- Very fast and simple method for quantitative release and recovery of biotinylated molecules
- Suitable for e.g. biotinylated proteins, peptides, DNA, RNA and PNA
- The method dissociates streptavidin – biotin complexes without affecting covalent bonds
- The elution substance SX is well-known, low priced (< 0.5$ per ml), save, and easy to handle
Due to the sublime performance with respect to simplicity, cost-effectiveness, and quantitative recovery of purified biotinylated molecules, the method is suitable for the elution of all sorts of biotin-streptavidin complexes.