Antibody targeted to tissue factor, preparation method therefor, and use thereof
The present invention provides a tissue factor (TF) monoclonal antibody and a preparation method therefor. The monoclonal antibody provided by the present invention can specifically bind with a TF antigen, has high affinity and low immunogenicity, and has the activity of resisting tumors and the like.
1. An anti-human tissue factor monoclonal antibody or an antigen-binding fragment thereof, wherein said antibody has
a heavy chain variable region comprising HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, and HCDR3 as set forth in SEQ ID NO: 3, and
a light chain variable region comprising LCDR1 as set forth in SEQ ID NO: 4, LCDR2 as set forth in SEQ ID NO: 5, and LCDR3 as set forth in SEQ ID NO: 6.
2. The antibody according to claim 1 , wherein the antibody is an animal-derived antibody, a chimeric antibody, or a humanized antibody.
3. The antibody according to claim 1 , wherein the heavy chain variable region sequence of the antibody is selected from the group consisting of SEQ ID NOS: 7, 9, 10, 11, 12, and 13; and/or
the light chain variable region sequence of the antibody is selected from the group consisting of SEQ ID NOS: 8, 14, 15, 16, and 17.
4. A method for treating cancer or a thrombotic disease, comprising administering a therapeutically effective amount of an anti-human tissue factor monoclonal antibody selected from the group consisting of TF-mAb-SC1, TF-mAb-Ch, TF-mAb-H29, TF-mAb-H30, TF-mAb-H31, TF-mAb-H32, TF-mAb-H33, TF-mAb-H34, TF-mAb-H35, TF-mAb-H36, TF-mAb-H37, TF-mAb-H38, TF-mAb-H39, TF-mAb-H40, TF-mAb-H41, TF-mAb-H42, TF-mAb-H43, TF-mAb-H44, TF-mAb-H45, TF-mAb-H46, TF-mAb-H47, and TF-mAb-H48 to a subject in need thereof.
5. The method according to claim 4 , wherein the method is for treating a thrombotic disease.
6. The method according to claim 4 , wherein the method is for treating cancer, and wherein the cancer is a cancer with high TF expression.
7. An isolated polynucleotide, which comprises a nucleic acid molecule encoding the heavy chain variable region of an anti-human tissue factor monoclonal antibody, and a nucleic acid molecule encoding the light chain variable region of said monoclonal antibody, wherein:
the heavy chain variable region of the antibody comprises HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, and HCDR3 as set forth in SEQ ID NO: 3, and
the light chain variable region of the antibody comprises LCDR1 as set forth in SEQ ID NO: 4, LCDR2 as set forth in SEQ ID NO: 5, and LCDR3 as set forth in SEQ ID NO: 6.
8. A vector, comprising the isolated polynucleotide according to claim 7 .
9. A genetically engineered host cell, comprising the vector according to claim 8 .
10. An immune cell, wherein the immune cell expresses the antibody according to claim 1 .
11. A pharmaceutical composition, comprising the antibody according to claim 1 and a pharmaceutically acceptable carrier.
12. A method for determining the presence or absence of a TF protein in a sample in vitro, the method comprises:
(1) contacting the sample with the antibody according to claim 1 in vitro; and
(2) determining whether an antigen-antibody complex is formed, wherein the formation of the complex indicates the presence of a TF protein in the sample.
13. A method for preparing an anti-human tissue factor monoclonal antibody, comprising:
(a) culturing the host cell according to claim 9 under conditions suitable for expressing an anti-human tissue factor monoclonal antibody; and
(b) separating the antibody from the culture.
14. The antibody according to claim 3 , wherein the antibody is selected from the group consisting of: TF-mAb-SC1, TF-mAb-Ch, TF-mAb-H29, TF-mAb-H30, TF-mAb-H31, TF-mAb-H32, TF-mAb-H33, TF-mAb-H34, TF-mAb-H35, TF-mAb-H36, TF-mAb-H37, TF-mAb-H38, TF-mAb-H39, TF-mAb-H40, TF-mAb-H41, TF-mAb-H42, TF-mAb-H43, TF-mAb-H44, TF-mAb-H45, TF-mAb-H46, TF-mAb-H47, and TF-mAb-H48.
15. The antibody according to claim 1 , wherein the antibody has an EC50 of 0.01-0.03 nM for the affinity to human TF protein.
16. The isolated polynucleotide according to claim 7 , wherein:
the heavy chain variable region sequence of the antibody is SEQ ID NO: 7, 9, 10, 11, 12, or 13; and/or
the light chain variable region sequence of the antibody is SEQ ID NO: 8, 14, 15, 16, or 17.
17. The isolated polynucleotide according to claim 7 , wherein the antibody is selected from the group consisting of: TF-mAb-SC1, TF-mAb-Ch, TF-mAb-H29, TF-mAb-H30, TF-mAb-H31, TF-mAb-H32, TF-mAb-H33, TF-mAb-H34, TF-mAb-H35, TF-mAb-H36, TF-mAb-H37, TF-mAb-H38, TF-mAb-H39, TF-mAb-H40, TF-mAb-H41, TF-mAb-H42, TF-mAb-H43, TF-mAb-H44, TF-mAb-H45, TF-mAb-H46, TF-mAb-H47, and TF-mAb-H48.