US 12,485,184 B2
Multimeric T-cell modulatory polypeptides and methods of use thereof
Saso Cemerski, Boston, MA (US); Ronald D. Seidel, III, Boston, MA (US); Rodolfo J. Chaparro, Cambridge, MA (US); and John F. Ross, Boston, MA (US)
Assigned to Cue Biopharma, Inc., Boston, MA (US)
Filed by Cue Biopharma, Inc., Boston, MA (US)
Filed on Nov. 16, 2023, as Appl. No. 18/511,502.
Application 18/511,502 is a division of application No. 18/334,911, filed on Jun. 14, 2023, granted, now 11,878,062.
Application 18/334,911 is a continuation of application No. 17/900,636, filed on Aug. 31, 2022.
Application 17/900,636 is a continuation of application No. PCT/US2021/031707, filed on May 11, 2021.
Claims priority of provisional application 63/041,451, filed on Jun. 19, 2020.
Claims priority of provisional application 63/023,834, filed on May 12, 2020.
Prior Publication US 2024/0189437 A1, Jun. 13, 2024
Int. Cl. G01N 33/53 (2006.01); A61K 39/00 (2006.01); A61K 47/64 (2017.01)
CPC A61K 47/646 (2017.08) [A61K 39/001114 (2018.08); A61K 39/001153 (2018.08); A61K 47/642 (2017.08); A61K 47/6425 (2017.08); C07K 2319/30 (2013.01)] 1 Claim
 
1. A method of detecting, in a mixed population of T cells obtained from an individual, the presence of a T cell that binds a Wilms tumor-1 (WT1) epitope presented by a peptide-major histocompatibility complex (pMHC) complex, the method comprising the steps of:
A) contacting in vitro the mixed population of T cells with a homodimer comprising two heterodimers, wherein each heterodimer comprises:
a) a first polypeptide having the amino acid sequence set forth in SEQ ID NO: 423 comprising:
i) a Wilms tumor-1 (WT1) peptide;
ii) a β2-microglobulin (β2M) polypeptide; and
iii) a peptide linker between the WT1 peptide and the β2M polypeptide, wherein the peptide linker comprises a Cys residue, and
b) a second polypeptide having the amino acid sequence set forth in SEQ ID NO: 486 comprising:
i) two variant IL-2 polypeptides;
ii) a major histocompatibility complex (MHC) class I heavy chain polypeptide; and
ii) an immunoglobulin (Ig) Fc polypeptide,
wherein the WT1 peptide, the β2M polypeptide, and the MHC class I heavy chain polypeptide form a pMHC complex,
wherein each heterodimer comprises a first disulfide bond formed between (i) the Cys residue in the peptide linker between the WT1 peptide and the β2M polypeptide, and (ii) a Cys residue in the MHC class I heavy chain polypeptide,
wherein each heterodimer comprises a second disulfide bond formed between a Cys residue in the β2M polypeptide and a Cys residue in the MHC class I heavy chain polypeptide, and
wherein two disulfide bonds link the Ig Fc polypeptide of one heterodimer to the Ig Fc polypeptide of the other heterodimer, and
B) detecting activation and/or proliferation of T cells comprising a T cell receptor that binds to the pMHC complex of each heterodimer.