Engineered Streptavidin Mutants with High-Affinities and Reversible Binding Capacities to Biotin- and SBP- Tagged Proteins


Description of Technology

The high-affinity interactions of biotin and streptavidin binding peptide (SBP) with streptavidin are well characterized and have been exploited for a variety of uses in biomedical research. One such application of this technology is the purification of recombinant biotinylated and SBP-tagged proteins via affinity chromatography. There are, however, several limitations that exist with respect to the technologies currently available in this field; wild-type streptavidin columns can only be used in a single round of purification due to the ultra-high affinity interaction between streptavidin and the free biotin in the elution buffers. The present technologies overcome these limitations.

Researchers at the University of Calgary have engineered novel streptavidin mutants that can be re-used in multiple rounds of biotin- and SBP-tagged protein purification. The mutant forms of streptavidin have a high binding affinity for SBP and a reduced binding affinity for biotin. This makes their interactions with biotin reversible and thus makes them viable for multiple rounds of purification. Moreover, they can easily be produced and manufactured using common bacterial expression systems and are readily purified.

Areas of Application
  • Affinity purification and biotinylated and SBP-tagged proteins using liquid chromatography
  • Immobilization of proteins to biochips, biosensors and enzyme bioreactors coated with the engineered streptavidin mutants
  • Conjugated mutants can serve as strippable detecting agents for various blots (i.e., Western blots)
  • Cell and protein isolation
  • Scalable up to 300mL
IP Status
Competitive Advantages
  • Efficiency affinity purification of biotin- and SBP- tagged proteins in a reusable and cost-effective manner - the engineered mutants can be regenerated for multiple rounds of protein purification
  • Mild elution conditions preserve the integrity of the molecule interest
  • Easily produced, purified and packaged into a product