A novel system with an improved efficiency for in vitro, as well as in vivo, biotinylation of a large amount of Avi-tagged target proteins. This system is an improvement over the only pre-existing commercial systems available for in vivo biotinylation because it greatly improves biotinylation efficiency and enables the complete labeling of target proteins. In addition, this MBP-BirA fusion encoding vector can be used stand-alone to produce a large amount of enzymatically active BirA biotin ligase that can be utilized for in vitro biotinylation.
Expanded Description:
UT Health San Antonio seeks to partner with a biological services company to develop and commercialize a novel system for in vitro as well as in vivo biotinylation for the successful labeling of proteins.
Proteins can be covalently labeled with biotin through chemical or enzymatic methods. Biotin ligases such as BirA can specifically biotinylate lysine side chain within an Avi-tag, which is a 15-amino acid acceptor peptide. The extremely tight binding between biotin and avidin or streptavidin makes biotin-labeling a very useful tool for many applications such as detection, immobilization and purification of proteins. Unfortunately there are very limited commercial kits in the market available only for in vitro biotinylation and furthermore these are not very cost-effective as they can only label very small amount of proteins.
Drs. Li and Sousa at the UT Health have developed a novel system with an improved efficiency for in vitro as well as in vivo biotinylation of large amount of Avi-tagged target proteins. Their system is an improvement over the only pre-existing commercial system available for in vivo biotinylation and addresses all their shortcomings. They have created a two-component kit, wherein the Avi-tagged target protein of interest is first highly expressed from an engineered vector. The second component comprises another vector encoding MBP-BirA biotin ligase fusion protein, which is expressed at high-level in soluble form. By co-expression of Avi-tagged target and the highly soluble MBP-BirA fusion, this system consequently greatly improves in vivo biotinylation efficiency and enables the complete labeling of target proteins. In addition, the MBP-BirA fusion encoding vector can be used standalone to produce large amount of enzymatically active BirA biotin ligase that can be utilized for in vitro biotinylation.
Commercial Applications & Advantages:
Biopharmaceutical companies and bio scientific researchers would be interested in the present invention.
- High efficiency of in vitro and in vivo biotinylation
- Complete labeling of proteins
- High expression and solubility of MBP-BirA biotin ligase fusion protein
- Novel method which enables relabeling of substrates in vitro
- Highly cost-effective
References: Biotechnol Lett. 2012 May 8. Novel system for in vivo biotinylation and its application to crab antimicrobial protein scygonadin. Li Y, Sousa R