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”