Antigen specific T-cell
Peptide MHC complex
T-cell receptor
HIV specific
HIV specific
HIV derived peptide-MHC
T-cell tagging strategy pioneered by Altman et al
Peptide MHC complex
MultimericBindingComplex
Antigen specific T-cell
T-cell tagging strategy pioneered by Altman et al
Antigen specific T-cell
CLAIM 12: said multimeric binding complex is formed by binding said monomer to a multivalent entity through specific attachment sites
Proimmune Pro5 Pentamer
Pentameric entity with 5 peptide-MHCcomplexes capable of binding toT-cell receptor molecules
Peptide-MHC complex
Multivalent coiled-coil domain
Site where pMHC monomer is specifically bound to the coiled-coil domain
Non-polar surfaces
The Pro5 coiled-coil domain is a multivalent entity
“Multivalency describes the binding of two (or more) entities that involves the simultaneous interaction between multiple, complementary functionalities on these entities”
Mulder et al, Org. and Biomol. Chem., (2004)
What holds the coiled-coil domain together?
Hydrogen bonds
Hydrophobic interactions
Acceptor
Donor
Multiple, simultaneous interactions betweencomplementary functionalities hold the helices together
Multimerization methods in ‘363 specification
(6:3-5) “Preferably, the multimer will be formed by binding the monomers to a multivalent entity through specific attachment sites”
(6:6-7) “The multimer may also be formed by chemical cross-linking of the monomers”
(6:20-23) “The attachment site for binding to a multivalent entity … will be a specific binding pair member or one that is modified to provide a specific binding pair member, where the complementary pair has a multiplicity of specific binding sites.”
(6:23-25) “Binding to the complementary binding member can be a chemical reaction, epitope-receptor binding, or hapten-receptor binding.
(6:58-59) “The multivalent binding partner may be free in solution, or may be attached to an insoluble support.”
Multivalency Self-multimerization
bindingepitope
receptor
specific binding pair
Coiled-coil from Pro5 Pentamer
Interactions span thelength of the helicesand exist between allmembers of the complex
There is no “specific binding pair”
There is no “multivalent binding partner”
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