MHC multimers, methods for their generation, labeling and use
The present invention relates to a soluble negative control MHC multimer comprising a nonsense peptide that binds the MHC protein efficiently, but that does not support binding of the resultant MHC-peptide complex to the desired T Cell Receptor. The nonsense peptide is designed to i) have a length enabling binding to the MHC allele in question, ii) have appropriate amino acids at relevant anchor positions which anchor the nonsense peptide to the peptide-binding groove of the MHC, iii) have amino acids outside the anchor positions that do not support binding to a T Cell Receptor, and iv) have an amino acid sequence that is different from the linear sequence of any naturally occurring peptide.
1. A soluble MHC multimer comprising (a-b-P) n , wherein n>1;
wherein a and b together form a functional MHC protein capable of binding nonsense peptide P;
wherein (a-b-P) is an MHC-peptide complex formed when P binds to the functional MHC protein;
wherein the functional MHC protein is HLA-A*0201:
wherein each MHC peptide complex of the multimer is associated with one or more multimerization domains;
and
wherein said nonsense peptide P is GLAGDVSAV (SEQ ID NO:11) or ALIAPVHAV (SEQ ID NO:12).
2. The soluble MHC multimer according to claim 1 , wherein the one or more multimerization domains comprise one or more polysaccharides.
3. The soluble MHC multimer according to claim 1 , wherein the one or more multimerization domains comprise one or more dextran moieties.
4. A composition comprising a plurality of soluble MHC multimers according to claim 1 , wherein the MHC multimers are identical or different, and a carrier.
5. The soluble MHC multimer according to claim 1 , wherein said one or more multimerization domains are soluble polysaccharides.
6. The soluble MHC multimer according to claim 5 , wherein said soluble polysaccharides are soluble dextrans.
7. The soluble MHC multimer according to claim 1 , wherein one or more labels are attached directly to the MHC multimer.
8. The soluble MHC multimer according to claim 1 , wherein one or more labels are attached indirectly to the MHC multimer via one or more marker molecules carrying one or more labels.