TGF-β3 specific antibodies and methods and uses thereof
Specific binding members, particularly antibodies and fragments thereof, which bind to transforming growth factor beta 3 (TGF-β3) are provided, particularly recognizing human and mouse TGF-β3, particularly antibodies and fragments that do not recognize or bind TGF-β1 or TGF-β2. Particular antibodies are provided which specifically recognize and neutralize TGF-β3. These antibodies are useful in the diagnosis and treatment of conditions associated with activated or elevated TGF-β3, including cancer, and for modulating immune cells and immune response, including immune response to cancer or cancer antigens. The anti-TGF-β3 antibodies, variable regions or CDR domain sequences thereof, and fragments thereof may also be used in therapy in combination with chemotherapeutics, immune modulators, or anti-cancer agents and/or with other antibodies or fragments thereof. Antibodies of this type are exemplified by the novel antibodies hereof, including antibody MTGF-β3-9, MTGF-β3-12, MTGF-β3-16, MTGF-β3-17 and MTGF-β3-19, whose sequences are provided herein.
1. An isolated antibody molecule or antigen binding fragment thereof which recognizes human and mouse transforming growth factor beta 3 (TGF-β3) and which neutralizes activity of TGF-β3, which is an antibody or fragment comprising:
(a) a light chain variable region comprising a CDR1 sequence KASQSVINDVA (SEQ ID NO:1) or KASQSVINAVA (SEQ ID NO:7), a CDR2 sequence YASNRYT (SEQ ID NO:2), and a CDR3 sequence QQDYSSPYT (SEQ ID NO:3), and a heavy chain variable region sequence comprising a CDR1 sequence SSWMH (SEQ ID NO:4) or SSWIH (SEQ ID NO:8), a CDR2 sequence RIFPGDGDTIYNGNFKG (SEQ ID NO:5) or RIYPGDGDTNYTGKFKG (SEQ ID NO:9), and a CDR3 sequence RMITTQAAMDY (SEQ ID NO:6);
(b) a light chain variable region comprising a CDR1 sequence KSSQSLLNSGNQKNYLA (SEQ ID NO:10), a CDR2 sequence GASTRES (SEQ ID NO:11), and a CDR3 sequence QNDHGFPLT (SEQ ID NO:12), and a heavy chain variable region sequence comprising a CDR1 sequence DYYIN (SEQ ID NO:13), a CDR2 sequence KIGPGTGRTYYNEKFKG (SEQ ID NO:14), and a CDR3 sequence YYGWGYAMDY (SEQ ID NO:15); or
(c) a light chain variable region comprising a CDR1 sequence RSSQSLIHSHGNTYLH (SEQ ID NO:16), a CDR2 sequence KLSNRFS (SEQ ID NO:17), and a CDR3 sequence SQSTHVPFT (SEQ ID NO:18), and a heavy chain variable region sequence comprising a CDR1 sequence SYWIT (SEQ ID NO:19), a CDR2 sequence DIFPGTGSTNYNEKFKT (SEQ ID NO:20), and a CDR3 sequence KLGPNYAVDY (SEQ ID NO:21).
2. The isolated antibody or fragment of claim 1 which does not react with TGF-β1 or TGF-β2.
3. The isolated antibody or fragment of claim 1 comprising:
(a) a light chain variable region comprising a CDR1 sequence KASQSVINDVA (SEQ ID NO:1), a CDR2 sequence YASNRYT (SEQ ID NO:2), and a CDR3 sequence QQDYSSPYT (SEQ ID NO:3), and a heavy chain variable region sequence comprising a CDR1 sequence SSWMH (SEQ ID NO:4), a CDR2 sequence RIFPGDGDTIYNGNFKG (SEQ ID NO:5), and a CDR3 sequence RMITTQAAMDY (SEQ ID NO:6);
(b) a light chain variable region comprising a CDR1 sequence KASQSVINAVA (SEQ ID NO:7), a CDR2 sequence YASNRYT (SEQ ID NO:2), and a CDR3 sequence QQDYSSPYT (SEQ ID NO:3), and a heavy chain variable region sequence comprising a CDR1 sequence SSWIH (SEQ ID NO:8), a CDR2 sequence RIYPGDGDTNYTGKFKG (SEQ ID NO:9), and a CDR3 sequence RMITTQAAMDY (SEQ ID NO:6); or
(c) a light chain variable region comprising a CDR1 sequence KSSQSLLNSGNQKNYLA (SEQ ID NO:10), a CDR2 sequence GASTRES (SEQ ID NO:11), and a CDR3 sequence QNDHGFPLT (SEQ ID NO:12), and a heavy chain variable region sequence comprising a CDR1 sequence DYYIN (SEQ ID NO:13), a CDR2 sequence KIGPGTGRTYYNEKFKG (SEQ ID NO:14), and a CDR3 sequence YYGWGYAMDY (SEQ ID NO:15).
4. The isolated antibody or fragment of claim 1 which is an antibody or antibody fragment comprising heavy chain variable region CDRs wherein the CDR1 domain comprises a sequence SSWXH wherein X is either M or I (SEQ ID NO:42), the CDR2 domain comprises a sequence RIFPGDGDTIYNGNFKG (SEQ ID NO:5) or RIYPGDGDTNYTGKFKG (SEQ ID NO:9) and the CDR3 domain comprises a sequence RMITTQAAMDY (SEQ ID NO:6).
5. The antibody or fragment of claim 4 further comprising light chain variable region CDRs wherein the CDR1 domain comprises a sequence KASQSVINXVA wherein X is either D or A (SEQ ID NO:44), the CDR2 domain comprises a sequence YASNRT (SEQ ID NO:2), and the CDR3 domain comprises a sequence QQDYSSPYT (SEQ ID NO:3).
6. The isolated antibody or fragment of claim 1 which is an antibody or antibody fragment comprising a heavy chain variable region amino acid sequence selected from the amino acid sequence set out in SEQ ID NO: 23, 27, 31, 35 or 39, or variants thereof having at least 90% amino acid identity to the heavy chain variable region sequence set out in SEQ ID NO: 23, 27, 31, 35 or 39, wherein said variants retain TGF-β3 reactivity and neutralization.
7. The isolated antibody or fragment of claim 6 further comprising a light chain variable region comprising an amino acid sequence selected from the amino acid sequence as set out in SEQ ID NO: 25, 29, 33, 37 or 41, or variants thereof having at least 90% amino acid identity to the light chain variable region sequence set out in SEQ ID NO: 25, 29, 33, 37 or 41, wherein said variants retain TGF-β3 reactivity and neutralization.
8. The isolated antibody of claim 6 which comprises a heavy chain having a variable region amino sequence comprising the amino acid sequence as set out in SEQ ID NO: 23, 31 or 39 or comprising the CDR domain sequences CDR1 sequence SSWMH (SEQ ID NO:4) or SSWIH (SEQ ID NO:8), CDR2 sequence RIFPGDGDTIYNGNFKG (SEQ ID NO:5) or RIYPGDGDTNYTGKFKG (SEQ ID NO:9), and CDR3 sequence RMITTQAAMDY (SEQ ID NO:6).
9. The isolated antibody of claim 7 which comprises a light chain having a variable region amino sequence comprising the amino acid sequence as set out in SEQ ID NO: 25, 33 or 41 or comprising the CDR region sequences CDR domain sequences CDR1 sequence KASQSVINDVA (SEQ ID NO:1) or KASQSVINAVA (SEQ ID NO:7), CDR2 sequence YASNRYT (SEQ ID NO:2), and CDR3 sequence QQDYSSPYT (SEQ ID NO:3).
10. The isolated antibody or fragment of claim 1 which is an antibody or fragment thereof wherein said isolated antibody is a F(ab′)2, scFv fragment, domain antibody, minibody, diabody, triabody or tetrabody.
11. The isolated antibody or fragment of claim 1 further comprising a detectable or functional label.
12. The isolated antibody of claim 11 , wherein said detectable or functional label is a covalently attached drug.
13. The isolated antibody of claim 11 , wherein said label is a radiolabel.
14. A kit for the diagnosis or prognosis of cancer in which TGF-β3 antigen is expressed, said kit comprising an antibody or fragment of claim 1 , with reagents and/or instructions for use.
15. A pharmaceutical composition comprising an antibody or fragment as defined in claim 1 and a pharmaceutically acceptable vehicle, carrier or diluent.
16. An immunological composition comprising an antibody or fragment as defined in claim 1 and a pharmaceutically acceptable vehicle, carrier or diluent, and further comprising one or more of an adjuvant, one or more antigen, an immunoregulatory antibody, or a small molecule inhibitor to an immune modulator.
17. A kit for the treatment of a tumor in a human patient, comprising a pharmaceutical dosage form of the pharmaceutical composition of claim 15 , and a separate pharmaceutical dosage form comprising an additional anti-cancer agent selected from the group consisting of cancer or tumor antigen(s), chemotherapeutic agents, radioimmunotherapeutic agents, and combinations thereof.
18. A method for detecting the presence and/or amount of TGF-β3 comprising:
(A) contacting a biological sample from a mammal with the antibody or fragment of claim 1 under conditions that allow binding of said TGF-β3 to said antibody to occur; and
(B) detecting whether binding has occurred between said TGF-β3 from said sample and the antibody, or determining the amount of binding that has occurred between said TGF-β3 from said sample and the antibody.