IP Library › Granted Patent US 12,590,152
Granted Patent B2
US 12,590,152 · App. 17/427,592 · Granted Mar 31, 2026

CD3-specific binding molecules

Inventors: Martina Canestraro (Abingdon, GB); Nele Dieckmann (Abingdon, GB); Stephen Harper (Abingdon, GB); Peter Benedict Kirk (Abingdon, GB); Rachel Mulvaney (Abingdon, GB); Ronan O'Dwyer (Abingdon, GB); Ian Butler Robertson (Abingdon, GB)
Assignee: IMMUNOCORE LIMITED
C07K16/2809A61K40/33C07K16/46C07K2317/24C07K2317/31C07K2317/565C07K2317/567C07K2317/622C07K2317/73C07K2317/92C12N15/00
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,590,152
App. No.
17/427,592
Granted
Mar 31, 2026
Kind
B2
Abstract

The present invention relates to specific binding molecules which bind to CD3, particularly antibodies and fragments thereof, with improved properties.

Claims (100)

1 . An antibody or antibody fragment thereof that specifically binds to CD3, comprising an immunoglobulin variable light chain (VL) domain and an immunoglobulin variable heavy chain (VH) domain, wherein the immunoglobin VL domain comprises the sequence:

(SEQ ID NO: 16)

AIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIY

YTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTF

GQGTKVEIK;

and

the immunoglobulin VH domain comprises the sequence:

(SEQ ID NO: 18)

EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYAMNWVRQAPGKGLEWVAL

INPYKGVSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSG

YYGDSDWYFDVWGQGTLVTVSS;

or

(SEQ ID NO: 19)

EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYAMNWVRQAPGKGLEWVAL

INPYKGVSTYNQKFKDRFTFSVDKSKNTAYLQMNSLRAEDTAVYYCARSG

YYGDSDWYFDVWGQGTLVTVSS.

2 . The antibody or antibody fragment thereof of claim 1 , comprising a single chain variable fragment (scFv) molecule.

3 . The antibody or antibody fragment thereof of claim 1 , wherein the immunoglobin VL domain and the immunoglobulin VH domain are connected via a linker.

4 . A fusion polypeptide comprising:

i) a targeting moiety selected from a T cell receptor (TCR), an antibody, or an antibody fragment; and

ii) the antibody or antibody fragment thereof of claim 1 .

5 . The fusion polypeptide of claim 4 , wherein the TCR is a heterodimeric alpha/beta TCR polypeptide pair or a single chain alpha/beta TCR polypeptide.

6 . The fusion polypeptide of claim 4 , wherein the TCR comprises a non-native disulfide bond between the constant region of the alpha chain and the constant region of the beta chain.

7 . The fusion polypeptide of claim 4 , wherein the antibody or antibody fragment thereof is fused to the C terminus or the N terminus of the targeting moiety, optionally via a linker.

8 . A pharmaceutical composition comprising the antibody or antibody fragment thereof of claim 1 .

9 . A nucleic acid molecule encoding the antibody or antibody fragment thereof of claim 1 .

10 . An expression vector comprising the nucleic acid molecule of claim 9 .

11 . An isolated host cell comprising the expression vector of claim 10 , wherein the nucleic acid encoding the antibody or antibody fragment thereof is present as a single open reading frame or two distinct open reading frames.

12 . A method of making an antibody or antigen binding fragment thereof that specifically binds to CD3, comprising culturing the isolated host cell of claim 11 under conditions for expression of a nucleic acid encoding the antibody or antibody fragment thereof and isolating the antibody or antibody fragment thereof.

13 . The antibody or antibody fragment thereof of claim 1 , wherein the immunoglobin VL domain comprises the sequence:

(SEQ ID NO: 16)

AIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIY

YTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTF

GQGTKVEIK;

and

the immunoglobin VH domain comprises the sequence:

EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYAMNWVRQAPGKGLEWVALINPYK GVSTYNQKFKDRFTFSVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDS DWYFDVWGQGTLVTVSS (SEQ ID NO: 19).

14 . The antibody or antibody fragment thereof of claim 2 , wherein the scFv molecule has the sequence:

(SEQ ID NO: 21)

AIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIY

YTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTF

GQGTKVEIKGGGGSGGGGSGGGGSGGGGSGGGSEVQLVESGGGLVQPGG

SLRLSCAASGYSFTGYAMNWVRQAPGKGLEWVALINPYKGVSTYNQKFK

DRFTFSVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQ

GTLVTVSS.

15 . The antibody or antibody fragment thereof of claim 1 , wherein the immunoglobin VL domain comprises the sequence:

(SEQ ID NO: 16)

AIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIY

YTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTF

GQGTKVEIK;

and

the immunoglobin VH domain comprises the sequence:

(SEQ ID NO: 18)

EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYAMNWVRQAPGKGLEWVA

LINPYKGVSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVYYCAR

SGYYGDSDWYFDVWGQGTLVTVSS.

16 . The antibody or antibody fragment thereof of claim 2 , wherein the scFv molecule has the sequence:

(SEQ ID NO: 20)

AIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIY

YTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTF

GQGTKVEIKGGGGSGGGGSGGGGSGGGGSGGGSEVQLVESGGGLVQPGG

SLRLSCAASGYSFTGYAMNWVRQAPGKGLEWVALINPYKGVSTYNQKFK

DRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQ

GTLVTVSS.

17 . An antibody or antigen binding fragment thereof that specifically binds to CD3 comprising an immunoglobulin VL domain comprising Complementarity Determining Regions (CDRs) VLCDR1, VLCDR2, and VLCDR3, and an immunoglobulin VH domain comprising CDRs VHCDR1, VHCDR2, and VHCDR3, wherein the amino acid sequence of;

VLCDR1 is 

(SEQ ID NO: 1)

QDIRNY;

VLCDR2 is 

YTS;

VLCDR3 is 

(SEQ ID NO: 2)

QQGNTLPWT;

VHCDR1 is 

(SEQ ID NO: 3)

GYSFTGYA;

VHCDR2 is 

(SEQ ID NO: 4)

INPYKGVS; and

VHCDR3 is 

(SEQ ID NO: 5)

ARSGYYGDSDWYFDV.

18 . A heterodimeric TCR-anti-CD3 antibody fusion molecule, comprising:

(a) a first polypeptide chain that comprises a TCR alpha chain variable domain; and

(b) a second polypeptide chain that comprises a TCR beta chain variable domain fused to an anti-CD3 scFv having the amino acid sequence:

(SEQ ID NO: 20) 

AIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIY

YTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTF

GQGTKVEIKGGGGSGGGGSGGGGSGGGGSGGGSEVQLVESGGGLVQPGG

SLRLSCAASGYSFTGYAMNWVRQAPGKGLEWVALINPYKGVSTYNQKFK

DRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQ

GTLVTVSS; or

(SEQ ID NO: 21)

AIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIY

YTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTF

GQGTKVEIKGGGGSGGGGSGGGGSGGGGSGGGSEVQLVESGGGLVQPGG

SLRLSCAASGYSFTGYAMNWVRQAPGKGLEWVALINPYKGVSTYNQKFK

DRFTFSVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQ

GTLVTVSS.

19 . The antibody or antibody fragment thereof of claim 3 , wherein the linker comprises between 5 amino acid residues to 30 amino acid residues.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 19, 2021
From: CANESTRARO, MARTINA; DIECKMANN, NELE; HARPER, STEPHEN; KIRK, PETER BENEDICT; MULVANEY, RACHEL; O'DWYER, RONAN; ROBERTSON, IAN BUTLER
To: IMMUNOCORE LTD.
Reel/Frame 058167/0101 →
Priority Claims (1)
GB 1901305 · Jan 30, 2019 · national
Continuity (1)
Related Publication 20220119527A1 · Apr 21, 2022
References Cited (105)
US 3773919A · Boswell et al. · 1973 [cited by applicant]
US 4485045A · Regen · 1984 [cited by applicant]
US 4544545A · Ryan et al. · 1985 [cited by applicant]
US 4619794A · Hauser · 1986 [cited by applicant]
US 5225539A · Winter · 1993 [cited by applicant]
US 5821337A · Carter et al. · 1998 [cited by applicant]
US 12065475B2 · Conroy · 2024 [cited by examiner]
US 12103971B2 · Dembek · 2024 [cited by examiner]
US 12134647B2 · Jaworski · 2024 [cited by examiner]
US 12195534B2 · Mai · 2025 [cited by examiner]
US 20070071675A1 · Wu et al. · 2007 [cited by applicant]
US 20150166661A1 · Chen et al. · 2015 [cited by applicant]
US 20230348595A1 · Chillakuri · 2023 [cited by examiner]
US 20240092859A1 · Conroy · 2024 [cited by examiner]
US 20240190969A1 · Chillakuri · 2024 [cited by examiner]
US 20240254228A1 · Mai · 2024 [cited by examiner]
US 20250179178A1 · Mai · 2025 [cited by examiner]
DE 3218121A1 · 1983 [cited by applicant]
EP 0052522A2 · 1982 [cited by applicant]
EP 0088046A2 · 1983 [cited by applicant]
EP 0036676B1 · 1984 [cited by applicant]
EP 0184187A2 · 1986 [cited by applicant]
EP 0058481B1 · 1986 [cited by applicant]
EP 0142541B1 · 1987 [cited by applicant]
EP 0143949B1 · 1988 [cited by applicant]
EP 0125023B1 · 1991 [cited by applicant]
EP 0120694B1 · 1993 [cited by applicant]
EP 0239400B1 · 1994 [cited by applicant]
GB 2188638A · 1987 [cited by applicant]
RU 2650868C2 · 2018 [cited by applicant]
WO WO1993011161A1 · 1993 [cited by applicant]
WO WO1994013804A1 · 1994 [cited by applicant]
WO WO1998039482A1 · 1998 [cited by applicant]
WO WO1999018129A1 · 1999 [cited by applicant]
WO WO2001062908A2 · 2001 [cited by applicant]
WO WO2001077342A1 · 2001 [cited by applicant]
WO WO2003020763A2 · 2003 [cited by applicant]
WO WO2004033685A1 · 2004 [cited by applicant]
WO WO2006000830A2 · 2006 [cited by applicant]
WO WO2010133828A1 · 2010 [cited by examiner]
WO WO2011001152A1 · 2011 [cited by applicant]
WO WO2014012085A2 · 2014 [cited by applicant]
WO WO2017109496A1 · 2017 [cited by applicant]
WO WO2017175006A1 · 2017 [cited by applicant]
WO WO2018234319A1 · 2018 [cited by examiner]
U.S. Appl. No. 18/901,016, Jaworski, Jakub. [cited by examiner]
U.S. Appl. No. 18/966,386, Mai, Nicole. [cited by examiner]
U.S. Appl. No. 18/004,644, Leonard, Sarah. [cited by examiner]
U.S. Appl. No. 18/004,644, filed Jan. 6, 2023, Leonard, Sarah. [cited by examiner]
U.S. Appl. No. 19/104,630, filed Feb. 18, 2025, Conroy, Paul. [cited by examiner]
Falconer, R.J., et al.(2011), Stabilization of a monoclonal antibody during purification and formulation by addition of basic amino acid excipients. J. Chem. Technol. Biotechnol., 86: 942-948. https://doi.org/10.1002/jc… [cited by examiner]
Lodish, et al. Molecular Cell Biology. 7th ed. 2012. W.H. Freeman and Co. Chapter 5. (Year: 2012). [cited by examiner]
Arnett KL, et al. Crystal structure of a human CD3-epsilon/delta dimer in complex with a UCHT1 single-chain antibody fragment. Proc Natl Acad Sci U S A. Nov. 16, 2004;101(46):16268-73. doi: 10.1073/pnas.0407359101. Epub… [cited by examiner]
Badri, H., et al., “Optimization of radiation dosing schedules for proneural glioblastoma,” [cited by applicant]
Baylot, V., et al., “TCTP Has a Crucial Role in the Different Stages of Prostate Cancer Malignant Progression,” Results and Problems in Cell Differentiation, 2017, vol. 64, pp. 255-261. [cited by applicant]
Colman, P.M., “Effects of amino acid sequence changes on antibody-antigen interactions,” Research in Immunology, vol. 145, Issue 1, 1994, pp. 33-36. [cited by applicant]
International Preliminary Report on Patentability, Chapter 1, Patent Cooperation Treaty Application No. PCT/EP2020/052316, Jul. 27, 2021, 9 pages. [cited by applicant]
International Search Report and Written Opinion, Patent Cooperation Treaty Application No. PCT/EP2020/052315, May 8, 2020, 14 pages. [cited by applicant]
Pan, Q., et al., “Blocking Neuropilin-1 Function Has an Additive Effect with Anti-VEGF to Inhibit Tumor Growth,” [cited by applicant]
Rudikoff, S. et al., “Single amino acid substitution altering antigen-binding specificity,” Proceedings of the National Academy of Sciences, Mar. 1982, vol. 79, No. 6, pp. 1979-1983. [cited by applicant]
Spiess, C., et al., “Alternative molecular formats and therapeutic applications for bispecific antibodies,” [cited by applicant]
Altschul et al., “Basic local alignment search tool,” [cited by applicant]
Altschul et al., “Gapped BLAST and PSI-BLAST: a new generation of protein database search programs,” [cited by applicant]
Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A., & Struhl, K. (Eds.), (1992), [cited by applicant]
Baeuerle et al., “Bispecific T-Cell Engaging Antibodies for Cancer Therapy,” [cited by applicant]
Barbas et al., “In vitro evolution of a neutralizing human antibody to human immunodeficiency virus type 1 to enhance affinity and broaden strain cross-reactivity,” [cited by applicant]
Bodanzsky, M. and Bodanzsky, A. (Eds), [cited by applicant]
Bragado et al., “Allelic polymorphism in the coding region of human TCR Cα gene and characterization of structural variability in the α chain constant domain,” [cited by applicant]
Brinkmann et al., “The making of bispecific antibodies,” Mabs, Feb.-Mar. 2017; 9(2): 182-212. [cited by applicant]
Chang et al., “Opportunities and challenges for TCR mimic antibodies in cancer therapy,” [cited by applicant]
Dahan, R. et al., “T-cell-receptor-like antibodies—generation, function and application,” Expert Reviews in Molecular Medicine, Feb. 24, 2012, vol. 14:e6, pp. 1-17. [cited by applicant]
Dennis et al., “Albumin Binding as a General Strategy for Improving the Pharmacokinetics of Proteins,” [cited by applicant]
Devereux, et al., “A comprehensive set of sequence analysis programs for the VAX,” Nucleic Acids Research, vol. 12, Issue 1, Part 1, Jan. 11, 1984, pp. 387-395. [cited by applicant]
Dozier et al., “Site-Specific PEGylation of Therapeutic Proteins,” [cited by applicant]
Epel et al., “A functional recombinant single-chain T cell receptor fragment capable of selectively targeting antigen-presenting cells,” Cancer Immunology, Immunotherapy, Nov. 2002, 51(10):565-73. [cited by applicant]
Eppstein et al., “Biological activity of liposome-encapsulated murine interferon gamma is mediated by a cell membrane receptor,” [cited by applicant]
Gram et al., “In vitro selection and affinity maturation of antibodies from a naive combinatorial immunoglobulin library,” [cited by applicant]
Hollinger, P. et al., “‘Diabodies’: small bivalent and bispecific antibody fragments,” [cited by applicant]
Hoo et al., “Characterization of a single-chain T-cell receptor expressed in [cited by applicant]
Husain et al., “Expanding the Boundaries of Biotherapeutics with Bispecific Antibodies,” [cited by applicant]
Huston et al., “Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in [cited by applicant]
Hwang et al., “Hepatic uptake and degradation of unilamellar sphingomyelin/cholesterol liposomes: a kinetic study,” [cited by applicant]
Jefferis, R., “Glycosylation as a strategy to improve antibody-based therapeutics,” [cited by applicant]
Jevsevar et al., “PEGylation of therapeutic proteins,” [cited by applicant]
Karlin and Altschul, “Applications and statistics for multiple high-scoring segments in molecular sequences,” [cited by applicant]
Karlin and Altschul, “Methods for assessing the statistical significance of molecular sequence features by using general scoring schemes,” [cited by applicant]
Lefranc et al., “IMGT™, the International ImMunoGene Tics Information System,” [cited by applicant]
Liddy, N. et al., “ Monoclonal TCR-redirected tumor cell killing”, [cited by applicant]
Myers, E. W. et al., “Optimal alignments in linear space,” Computer applications in the biosciences (CABIOS), Mar. 1988, vol. 4, No. 1, pp. 11-17. [cited by applicant]
Pearson and Lipman, “Improved tools for biological sequence comparison,” PNAS, Apr. 1988, 85 (8) 2444-2448; https://doi.org/10.1073/pnas.85.8.2444. [cited by applicant]
Plückthun, “Antibody Engineering: Advances from the Use of [cited by applicant]
Reff, M. E., “High-level production of recombinant immunoglobulins in mammalian cells,” [cited by applicant]
Schellenberger et al., “A recombinant polypeptide extends the in vivo half-life of peptides and proteins in a tunable manner,” [cited by applicant]
Schier et al., “Isolation of Picomolar Affinity Anti-c-erbB-2 Single-chain Fv by Molecular Evolution of the Complementarity Determining Regions in the Center of the Antibody Binding Site,” [cited by applicant]
Schlapschy, M. et al., “PASylation: a biological alternative to PEGylation for extending the plasma half-life of pharmaceutically active proteins,” Protein Engineering, Design and Selection, vol. 26, Issue 8, Aug. 2013,… [cited by applicant]
Schodin, B. A. et al., “Binding properties and solubility of single-chain T cell receptors expressed in [cited by applicant]
Shalaby et al., “Development of humanized bispecific antibodies reactive with cytotoxic lymphocytes and tumor cells overexpressing the HER2 protooncogene,” [cited by applicant]
Sinclair, A.M. and Elliott, S., “Glycoengineering: The effect of glycosylation on the properties of therapeutic proteins,” [cited by applicant]
Torelli and Robotti, “Advance and ADAM: two algorithms for the analysis of global similarity between homologous informational sequences,” [cited by applicant]
Traunecker et al., “Bispecific single chain molecules (Janusins) target cytotoxic lymphocytes on HIV infected cells,” [cited by applicant]
Trill et al., “Production of monoclonal antibodies in COS and CHO cells,” [cited by applicant]
Ward, E.S. et al., “Binding activities of a repertoire of single immunoglobulin variable domains secreted from [cited by applicant]
Weidanz et al., “Display of functional αß single-chain T-cell receptor molecules on the surface of bacteriophage,” [cited by applicant]
Yuraszeck et al., “Translation and Clinical Development of Bispecific T-cell Engaging Antibodies for Cancer Treatment,” [cited by applicant]
Yaojian, C. et al., “Preparation and preliminary functional study of anti-EGFR/CD3 bispecific antibody,” Journal of Fujian Medical University, Issue 6, 2017, pp. 1-13 [Online] [Retrieved on Aug. 26, 2025] Retrieved from… [cited by applicant]