Promab Biotechnologies, Inc.

Forevertek Biotechnology Co., Ltd

Caribou Biosciences, Inc.

The General Hospital Corporation

Zhejiang Ruijiamei Biotech Co., Ltd.

Vita Golubovskaya

Lijun Wu

Marcela V Maus

Hua Zhou

Martyn Lewis

The present invention is directed to a monoclonal anti-human CS1 antibody or a single-chain variable fragment (scFv), comprising VH having the amino acid of SEQ ID NO: 3 and VL having the amino acid of SEQ ID NO: 7. The present invention is also directed to a chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) of the present invention, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain.

CS1 antibody and anti-CS1-CAR-t cells

The present invention is directed to a humanized BCMA single-chain variable fragment (scFv), comprising Vhaving the amino acid sequence of SEQ ID NO: 3 and Vhaving the amino acid sequence of SEQ ID NO: 5. The present invention is also directed to a BCMA chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) of the present invention, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain. This humanized BCMA-CAR-T cells have specific killing activity against BCMA-positive tumor cells.

Humanized BCMA antibody and BCMA-CAR-T cells

The present invention is directed to a humanized monoclonal anti-human EpCAM antibody, such as a single-chain variable fragment (scFv), comprising Vhaving the amino acid of SEQ ID NO: 2 and Vhaving the amino acid of SEQ ID NO: 4. The present invention is also directed to a chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) of the present invention, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain.

Epcam antibody and Epcam-CAR-T cells

Provided is a humanized CD19 single-chain variable fragment (scFv) having a mutation of valine to glycine in CDR1, comprising VH having the amino acid sequence of SEQ ID NO: 6 and VL having the amino acid sequence of SEQ ID NO: 5. Also provided is a CD19 chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv), (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain. This humanized CD19-CAR-T cells have specific killing activity with secretion of cytokine IFN-gamma in CAR-T cells in vitro and in vivo.

CAR-T cells with humanized CD19 ScFv with mutation in CDR 1 region

The present invention is directed to a humanized BCMA antibody or an antigen-binding fragment thereof, comprising Vhaving the amino acid sequence of SEQ ID NO: 3 and Vhaving the amino acid sequence of SEQ ID NO: 5. The present invention is also directed to a BCMA chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) of the present invention, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain. This humanized BCMA-CAR-T cells have specific killing activity against BCMA-positive tumor cells.

The present invention is directed to a humanized CD19 single-chain variable fragment (scFv), comprising the amino acid sequence of SEQ ID NO: 8. The present invention is also directed to a CD19 chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) of the present invention, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain. The humanized anti-CD19 scFv of the present invention exhibits selective and high-affinity binding to CD19. CD19-CAR T cells based on humanized scFv of the present invention are useful to treat patients with B-cell malignancies including leukemia and lymphomas.

CAR-T cells with humanized CD19 ScFv

The present invention is directed to a monoclonal anti-human BCMA antibody, or a single-chain variable fragment (scFv), comprising Vhaving the amino acid sequence of SEQ ID NO: 6 and Vhaving the amino acid sequence of SEQ ID NO: 7. The present invention is also directed to a BCMA chimeric antigen receptor (CAR) comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) of the present invention, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain. The monoclonal antibody of the present invention exhibits selective and high-affinity binding to BCMA. BCMA CAR-T cells based on BCMA scFv of the present invention significantly decreases multiple myeloma tumor growth in an animal model.

BCMA-CAR-T cells

The present invention is directed to a chimeric antigen receptor (CAR) fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) comprising Vand V, wherein scFv binds to human PLAP (placental alkaline phosphatase), (ii) a transmembrane domain, (iii) a co-stimulatory domain of CD28, OX-40, GITR, or 4-1BB, and (iv) CD3 an activating domain. The present invention is also directed to T cells, natural killer (NK) cells, or macrophages, modified to express the CAR of the present invention. The present invention is further directed to a method for treating PLAP-positive cancer cells by administering PLAP-CAR-T cells, PLAP-CAR-NK cells, or PLAP-CAR-macrophages to the patients.

PLAP-CAR-effector cells

The present invention is directed to a monoclonal mouse or humanized ROR1 antibody, or a single-chain variable fragment (scFv). The present invention is also directed to a mouse or humanized ROR1 chimeric antigen receptor (CAR) comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) of the present invention, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain.

Anti-ROR1 antibody and ROR1-targeting engineered cells

The present invention is directed to a chimeric antigen receptor fusion protein comprising: (i) a single-chain variable fragment (scFv) comprising Vand V, wherein scFv has an activity against a tumor antigen, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain; wherein the CAR further comprises a human transferrin fragment, which is an epitope for an antibody against human transferrin, at N-terminus or C-terminus to scFv, or between Vand V. Preferred tumor antigens are CD19, CD22 and BCMA. The CD19-TF-CAR-T cells, CD22-TF-CAR-T cells, and BAMA-TF CAR-T cells secrete less cytokines, but they have the same efficacy against cancer target cells when comparing with same CAR without TF.

Chimeric antigen receptors comprising a human transferrin epitope sequence

The present invention provides a chimeric antigen receptor (CAR) fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) comprising Vand V, wherein scFv has an activity against CD47, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain. In one embodiment, the scFv is derived from a humanized anti-CD47 antibody. The present invention also provides T cells modified to express the CAR of the present invention.

CD47-CAR-T cells

The present invention is directed to a chimeric antigen receptor (CAR) fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) comprising Vand V, wherein scFv binds to a tumor antigen, (ii) a transmembrane domain, (iii) a co-stimulatory domain of GITR intracellular domain, and (iv) an activating domain. In one embodiment, the tumor antigen is human epidermal growth factor receptor (EGFR), human mesothelin, or human CD19. CARs having GITR intracellular domain as a co-stimulatory domain have certain advantages over other traditional CAR co-stimulatory domains.

Growing community of inventors

Richmond, CA, United States of America

Vita Golubovskaya