# US 12,595,304 B2 · Antibodies specific to glycosylated LAG3 and methods of use thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,595,304 B2 - Document ID: US12595304B2 - Status: Granted - Kind: B2 - Application number: 17/767,894 - Filed: 2020-10-09 - Published: 2022-11-10 - Granted: 2026-04-07 - Earliest claimed: 2019-10-09 - Assignees: Stcube, Inc. - Applicants: Stcube & Co. - Inventors: Stephen S. Yoo - Web page: https://patentgrep.com/patent/US12595304B2 ## CPC - C07K 16/2803: {against the immunoglobulin superfamily} - A61P 35/00: Antineoplastic agents - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/33: Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity - C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues - C07K 2317/41: Glycosylation, sialylation, or fucosylation - C07K 2317/622: Single chain antibody (scFv) - C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract Antibodies that selectively bind to glycosylated LAG3 relative to unglycosylated LAG3 are provided. In some aspects, LAG3 polypeptides comprising glycosylated amino acid positions are also provided. Methods for making and using such antibodies and polypeptides (e.g., for the treatment of cancer) are also provided. ## Claims ### Claim 1 An isolated monoclonal antibody which selectively binds to glycosylated LAG3 relative to unglycosylated form of LAG3, said isolated monoclonal antibody having (I) a VH domain comprising (a) a Chothia CDR H1 with the amino acid sequence of SEQ ID NO: 6, a CDR H2 with the amino acid sequence of SEQ ID NO: 7, and a CDR H3 with the amino acid sequence of SEQ ID NO: 8 (b) an AbM CDR H1 with the amino acid sequence of SEQ ID NO: 9, a CDR H2 with the amino acid sequence of SEQ ID NO: 10, and a CDR H3 with the amino acid sequence of SEQ ID NO: 8; or (c) a Kabat CDR H1 with the amino acid sequence of SEQ ID NO: 11, a CDR H2 with the amino acid sequence of SEQ ID NO: 12, and a CDR H3 with the amino acid sequence of SEQ ID NO: 8; and (II) a VL domain comprising a CDR L1 with the amino acid sequence of SEQ ID NO: 16, a CDR L2 with the amino acid sequence of SEQ ID NO: 17, and a CDR L3 with the amino acid sequence of SEQ ID NO: 18, or said isolated monoclonal antibody having (I) a VH domain comprising a Contact CDR H1 with the amino acid sequence of SEQ ID NO: 13, a CDR H2 with the amino acid sequence of SEQ ID NO: 14, and a CDR H3 with the amino acid sequence of SEQ ID NO: 15 and (II) a VL domain comprising a Contact CDR L1 with the amino acid sequence of SEQ ID NO: 19, a CDR L2 with the amino acid sequence of SEQ ID NO: 20, and a CDR L3 with the amino acid sequence of SEQ ID NO: 21. ### Claim 2 (depends on 1) The isolated antibody of claim 1, wherein the antibody increases secretion of IFN-γ and/or IL-2 or wherein the antibody increases T-cell proliferation. ### Claim 3 (depends on 1) The isolated antibody of claim 1, wherein the antibody selectively binds to LAG3 glycosylated at positions N188, N250, N256, N343, or any combination thereof, relative to unglycosylated LAG3 and wherein the antibody blocks binding of LAG3 to one or more of Gal-3, MHCII, LSECtin, and CD3 and wherein the binding affinity of anti-glycLAG3 antibodies for glycosylated LAG3 is from 0.1-10 nM inclusive of the lower and upper values. ### Claim 4 (depends on 1) The isolated antibody of claim 1, wherein the VH domain has an amino acid sequence of SEQ ID NO: 3 and the VL has an amino acid sequence of SEQ ID NO: 5. ### Claim 5 (depends on 1) The isolated antibody of claim 1, wherein the VH domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 3. ### Claim 6 (depends on 1) The isolated antibody of claim 1, wherein the VL domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 5. ### Claim 7 (depends on 1) The isolated antibody of claim 1, which has a VH domain comprising a Chothia CDR H1 with an amino acid sequence of SEQ ID NO: 6, a CDR H2 with an amino acid sequence of SEQ ID NO: 7, and a CDR H3 with an amino acid sequence of SEQ ID NO: 8 or which has a VH domain comprising a AbM CDR H1 with an amino acid sequence of SEQ ID NO: 9, a CDR H2 with an amino acid sequence of SEQ ID NO: 10, and a CDR H3 with an amino acid sequence of SEQ ID NO: 8 or which has a VH domain comprising a Kabat CDR H1 with an amino acid sequence of SEQ ID NO: 11, a CDR H2 with an amino acid sequence of SEQ ID NO: 12, and a CDR H3 with an amino acid sequence of SEQ ID NO: 8 and which has a VL domain comprising a CDR L1 with an amino acid sequence of SEQ ID NO: 16, a CDR L2 with an amino acid sequence of SEQ ID NO: 17, and a CDR L3 with an amino acid sequence of SEQ ID NO: 18. ### Claim 8 (depends on 1) The isolated antibody of claim 1, which has a VH domain comprising a Contact CDR H1 with an amino acid sequence of SEQ ID NO: 13, a CDR H2 with an amino acid sequence of SEQ ID NO: 14, and a CDR H3 with an amino acid sequence of SEQ ID NO: 15 and which has a VL domain comprising a Contact CDR L1 with an amino acid sequence of SEQ ID NO: 19, a CDR L2 with an amino acid sequence of SEQ ID NO: 20, and a CDR L3 with an amino acid sequence of SEQ ID NO: 21. ### Claim 9 (depends on 1) The isolated antibody of claim 1, which has human framework regions. ### Claim 10 (depends on 1) The isolated antibody of claim 1, which comprises a human constant domain. ### Claim 11 (depends on 1) The isolated antibody of claim 1, wherein the antibody is an IgG, IgM, IgA or an antigen binding fragment thereof. ### Claim 12 (depends on 1) The isolated antibody of claim 1, wherein the antibody is a Fab′, a F(ab′)2, a F(ab′)3, a monovalent scFv, or a bivalent scFv. ### Claim 13 (depends on 1) The isolated antibody of claim 1, wherein the antibody is a human or humanized antibody. ### Claim 14 (depends on 1) The isolated antibody of claim 1, wherein the antibody is conjugated to an imaging agent, a chemotherapeutic agent, a toxin or a radionuclide. ### Claim 15 (depends on 1) A composition comprising the isolated antibody according to claim 1 in a pharmaceutically acceptable carrier. ### Claim 16 (depends on 1) A method for treating a subject having a cancer comprising administering an effective amount of an isolated antibody of claim 1 in a pharmaceutically acceptable composition to the subject, wherein the cancer is a breast cancer, lung cancer, head & neck cancer, prostate cancer, esophageal cancer, tracheal cancer, brain cancer, liver cancer, bladder cancer, stomach cancer, pancreatic cancer, ovarian cancer, uterine cancer, cervical cancer, testicular cancer, colon cancer, rectal cancer or skin cancer. ### Claim 17 (depends on 16) The method of claim 16, further comprising administering at least a second anti-cancer therapy to the subject, wherein the second anti-cancer therapy is a surgical therapy, chemotherapy, radiation therapy, cryotherapy, hormonal therapy, immunotherapy or cytokine therapy. ### Claim 18 (depends on 17) The method of claim 17, wherein the second anticancer therapy is durvalumab, nivolumab, pembrolizumab, avelumab, atezolizumab, cemiplimab, or ipilimumab. ### Claim 19 (depends on 1) A method for assessing LAG3 glycosylation comprising contacting a LAG3-containing sample with an antibody according to claim 1.