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Protein Engineering Technology Yields New Therapeutics

Technology Application
Purification of antigen-specific T cells, Allergy,Oncology, Auto-immunity, Infectious disease
Detailed Technology Description
New expression method significantly reduces the cost andcomplications associated with HLA/peptide production for use in the biotechsector
*Abstract

The development of fluorochrome-conjugated peptide-majorhistocompatibility complex (pMHC) multimers has transformed the study ofantigen-specific T cells by enabling their characterization and isolation fromex vivo samples. However, pMHCs productions are currently hindered becauselarge-scale manufacture is not possible and lacks critical proteinmodifications. A novelfluorochrome-conjugated human pMHC dimer with the advantage of reducingproduction time from fourteen days to two days, test tube peptide exchange,robust staining, and increased stability due to correct proteinmodification. Peptides have beenexchanged into the system and tested for specific recognition.  The novel technology is applicable in manyhuman disease applications including allergy, infectious disease, autoimmunedisorders, oncology, and protein therapeutics.


HLA class I molecules are heterodimers consisting oftwo polypeptide chains, α and b2-microglobulin(b2m), which arenon-covalently linked. While TCR on the surface of cytotoxic T cells recognizesthe peptides presented by the HLA. Therefore, enhancement of the CD8 and HLA class I interaction leads tothe improvement in the strength of the interaction. Improved methods in pMHCs production will lead to new therapeutics in manyfields (Nepom, 2012).

*Principal Investigator

Dr. Naoto Hirano, Princess Margaret CancerCentre, University Health Network

*Publications
Paul Klenerman, VincenzoCerundolo & P. Rod Dunbar. Tracking Tcells with tetramers: new tales from new tools. Nature Reviews Immunology2, 263-272 (2002)Stuart Sims, Christian Willberg& Paul Klenerman. MHC–peptide tetramers for the analysis ofantigen-specific T cells. Expert Review of Vaccines, 9:7, 765-774 (2010)Gerard Nepom. MHC Class II Tetramers. J Immunol, 188:2477-2482 (2012)
Country/Region
USA

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