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CN-118791604-B - Anti-Ki 67 antibody, antibody conjugate, kit, nucleic acid, cell and application thereof

CN118791604BCN 118791604 BCN118791604 BCN 118791604BCN-118791604-B

Abstract

The invention discloses an anti-Ki 67 antibody, an antibody conjugate, a kit, nucleic acid, cells and application thereof, and relates to the technical field of antibody preparation. According to the invention, the antibody specifically binding to the Ki67 protein is obtained through screening, and the antibody has better binding activity and affinity with the Ki67 protein through detection, so that a detection product based on a Ki67 protein marker can be developed. In addition, the anti-Ki 67 antibody or the antigen binding fragment thereof can be used for early warning, diagnosis, efficacy evaluation and prognosis of diseases using Ki67 protein as a marker, such as tonsil cancer and the like, is beneficial to early warning, screening, intervention and efficacy evaluation of tumor diseases, and has wide application prospect.

Inventors

  • FU KANG
  • YAN ZHENG
  • HU JUNAN

Assignees

  • 生工生物工程（上海）股份有限公司

Dates

Publication Date
20260512
Application Date
20240826

Claims (20)

  1. 1. An anti-Ki 67 antibody or antigen-binding fragment thereof, comprising the complementarity determining regions: HCDR1, which consists of the amino acid sequence shown in SEQ ID NO. 1; HCDR2, which consists of the amino acid sequence shown in SEQ ID NO. 2; HCDR3, which consists of the amino acid sequence shown in SEQ ID NO. 3; LCDR1, which consists of the amino acid sequence shown in SEQ ID NO. 4; LCDR2, which consists of the amino acid sequence shown in SEQ ID NO. 5; And LCDR3, which consists of the amino acid sequence shown in SEQ ID NO. 6.
  2. 2. The anti-Ki 67 antibody or antigen-binding fragment thereof of claim 1, further comprising a framework region of: HFR1, amino acid sequence having at least 80% homology with SEQ ID NO. 7; HFR2 has at least 80% homology with the amino acid sequence shown in SEQ ID NO. 8; HFR3 has at least 80% homology with the amino acid sequence shown in SEQ ID NO. 9; HFR4, amino acid sequence having at least 80% homology with SEQ ID NO. 10; LFR1, the amino acid sequence has at least 80% homology with the sequence shown in SEQ ID NO. 11; LFR2, amino acid sequence has at least 80% homology with SEQ ID NO. 12; LFR3 has at least 80% homology with the amino acid sequence shown in SEQ ID NO. 13; LFR4 has an amino acid sequence having at least 80% homology with the sequence shown in SEQ ID NO. 14.
  3. 3. The anti-Ki 67 antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof further comprises a constant region.
  4. 4. The anti-Ki 67 antibody or antigen binding fragment thereof of claim 3, wherein the constant regions comprise a heavy chain constant region and a light chain constant region.
  5. 5. The antibody or antigen-binding fragment thereof of claim 4, wherein the heavy chain constant region is selected from the group consisting of an IgG1, igG2, igG3, igG4, igA, igM, igE, and IgD heavy chain constant region and the light chain constant region is selected from the group consisting of kappa-type and lambda-type light chain constant regions.
  6. 6. The anti-Ki 67 antibody or antigen binding fragment thereof of claim 5, wherein the heavy chain constant region sequence has at least 80% homology to the sequence set forth in SEQ ID No. 15 and the light chain constant region sequence has at least 80% homology to the sequence set forth in SEQ ID No. 16.
  7. 7. The anti-Ki 67 antibody or antigen binding fragment thereof of claim 6, wherein the heavy chain constant region sequence is set forth in SEQ ID No. 15 and the light chain constant region sequence is set forth in SEQ ID No. 16.
  8. 8. The antibody or antigen-binding fragment thereof of claim 3, wherein the constant region is of a species origin of bovine, equine, porcine, ovine, caprine, rat, mouse, canine, feline, rabbit, donkey, deer, mink, chicken, duck, goose, or human.
  9. 9. The anti-Ki 67 antibody or antigen binding fragment thereof of claim 8, wherein the constant region is of murine or human species origin.
  10. 10. The anti-Ki 67 antibody or antigen-binding fragment thereof of any one of claims 1-9, wherein the antigen-binding fragment is selected from any one of F (ab ') 2, fab', fab, fv, and scFv of the antibody.
  11. 11. An antibody conjugate comprising the antibody or antigen-binding fragment thereof of any one of claims 1-10, wherein the antibody or antigen-binding fragment thereof is labeled with a detectable label.
  12. 12. The antibody conjugate of claim 11, wherein the detectable label is selected from the group consisting of a fluorescent dye, an enzyme that catalyzes the development of a substrate, a radioisotope, a chemiluminescent reagent, and a nanoparticle-based label, and wherein the antibody or antigen binding fragment thereof is coated on a solid phase.
  13. 13. The antibody conjugate of claim 12, wherein the solid phase is selected from the group consisting of microspheres, plates, and membranes.
  14. 14. The antibody conjugate of claim 12, wherein the solid phase is selected from the group consisting of magnetic microspheres, plastic microparticles, latex microspheres, microwell plates, glass, capillaries, nylon, and nitrocellulose membranes.
  15. 15. Use of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 10 or an antibody conjugate according to any one of claims 11 to 14 for the preparation of a product selected from the group consisting of a reagent, a kit or a chip; the product has at least one of Ki67 protein detection, ki67 protein enrichment and Ki67 protein purification.
  16. 16. The use according to claim 15, wherein said Ki67 protein detection is labelling of tissue cells with said antibody or antigen binding fragment thereof.
  17. 17. A reagent or kit for Ki67 protein detection, ki67 protein enrichment or Ki67 protein purification, comprising an antibody or antigen binding fragment thereof according to any one of claims 1-10 or an antibody conjugate according to any one of claims 11-14.
  18. 18. A nucleic acid encoding the antibody or antigen-binding fragment thereof of any one of claims 1-10.
  19. 19. A vector comprising the nucleic acid of claim 18.
  20. 20. A recombinant cell comprising the vector of claim 19.

Description

Anti-Ki 67 antibody, antibody conjugate, kit, nucleic acid, cell and application thereof Technical Field The invention relates to the technical field of antibody preparation, in particular to an anti-Ki 67 antibody, an antibody conjugate, a kit, nucleic acid, cells and application thereof. Background Clinically Ki67 is mainly used to label cells in the proliferation cycle, which is mainly due to the fact that the faster the tumor grows, the worse the tissue differentiation ability, and the more sensitive to Ki 67. Generally, such tumors have a poor prognosis, and are often difficult to cure completely. So that late stage tumors have increased sensitivity to Ki67 due to their faster division and less complete cell differentiation. The Ki-67 antigen is a protein related to cell nuclear division proliferation, is expressed in the cell cycle S, G, the G2 and the M phase, and is deleted in the G0 phase, so that the Ki-67 antigen is often used as a reliable marker of tumor cell proliferation activity. Clinical detection of protein expression in tumor cells is often performed by Immunohistochemical (IHC) pathology experiments, and the accuracy and sensitivity of the IHC experiments are determined by the quality of monoclonal antibodies that specifically bind the protein. Therefore, developing a monoclonal antibody with higher binding specificity aiming at Ki67 polypeptide has important significance for detecting the expression level of Ki67 protein. In view of this, the present invention has been made. Disclosure of Invention The invention aims to provide an anti-Ki 67 antibody, an antibody conjugate, a kit, nucleic acid, cells and application thereof, which are used for solving the technical problems. The invention is realized in the following way: in a first aspect, the invention provides an antibody or antigen binding fragment thereof against Ki67, the antibody comprising the following complementarity determining regions: HCDR1, which consists of the amino acid sequence shown in SEQ ID NO. 1; HCDR2, which consists of the amino acid sequence shown in SEQ ID NO. 2; HCDR3, which consists of the amino acid sequence shown in SEQ ID NO. 3; LCDR1, which consists of the amino acid sequence shown in SEQ ID NO. 4; LCDR2, which consists of the amino acid sequence shown in SEQ ID NO. 5; And LCDR3, which consists of the amino acid sequence shown in SEQ ID NO. 6. In a second aspect, the invention provides an antibody conjugate comprising an antibody or antigen binding fragment thereof as described above. In a third aspect, the invention provides the use of an antibody as defined above, or an antigen binding fragment thereof, or an antibody conjugate as defined above, in the preparation of any one of the following products, selected from reagents, kits or chips. In a fourth aspect, the invention provides a reagent or kit for tumor risk pre-warning, diagnosis, efficacy monitoring, prognosis evaluation, ki67 protein detection, ki67 protein enrichment or Ki67 protein purification, which contains the above antibody or antigen binding fragment thereof or the above antibody conjugate, and the tumor uses the Ki67 protein as a detection marker. In a fifth aspect, the invention provides a nucleic acid encoding an antibody or antigen binding fragment thereof as described above. In a sixth aspect, the invention provides a vector comprising a nucleic acid as described above. In a seventh aspect, the present invention provides a recombinant cell comprising the vector described above. The invention has the following beneficial effects: According to the invention, the antibody which specifically binds to the Ki67 protein is obtained through screening, and the antibody has better binding activity and affinity with the Ki67 protein through detection, so that detection products based on the Ki67 protein marker, such as detection reagents, kits or chips, can be developed. Detection of Ki67 using the anti-Ki 67 antibodies or antigen binding fragments thereof of the present invention can improve the sensitivity and specificity of the detection. In addition, the anti-Ki 67 antibody or antigen binding fragment thereof can be used for early warning, diagnosis, efficacy evaluation and prognosis of diseases with Ki67 protein as a marker, such as tonsil cancer and the like, and is beneficial to early warning, screening, intervention and efficacy evaluation of tumor diseases. Therefore, the invention provides more protein choices for detection of Ki67 protein, early warning, diagnosis, efficacy evaluation and prognosis of epithelial tumors. Drawings In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to t