Systems And Methods For The Preparation Of Peptide Receptive Mhc-I/Chaperone Complexes With Native Glycan Modifications

Background

Typically, peptide receptive MHC-I multimer reagents are prepared in bacterial (E. coli) culture. While this is efficient, it does not result in glycosylation of the MHC-I peptide fragments as is done in mammalian cells. As a result, if such reagents are produced in mammalian cells, proper glycosylation would result and the reagents would have a potentially more accurate representation of the natural T-cell target. 

Technology Description

This technology involves the coexpression of leucine zipper tagged single chain class I MHC molecules and the TAPBPR chaperone in mammalian cells to produce glycosylated MHC-I that are ready to accept an antigenic peptide of interest.

The expressed MHC-I can be purified from the supernatant, the leucine zippers removed through use of a specific protease (with a site engineered into the construct), multimerized, and contacted with the peptide of interest. In contrast with other technologies in this portfolio (e.g. 2018-408), no placeholder peptide is needed to create the peptide receptive MHC-I. 

Applications

Peptide-receptive (empty) MHC-I reagents 

MHC-I Multimer reagents

T Cell receptor discovery 

T Cell epitope identification 

Advantages

Glycosylated MHC-I are more realistic

Efficient production of soluble MHC-I reagents from mammalian cells

MHC-I can be purified from supernatants

No need for placeholder peptide 

Intellectual Property Information

Additional Patent Pending

Related Materials

• Production of soluble pMHC-I molecules in mammalian cells using the molecular chaperone TAPBPR - 12/31/2019