The preparation of VEGFR1/CD3 bispecific antibody and its specific cytotoxicity against VEGFR1-positive breast cancer cells

Biotechnol Appl Biochem. 2014 Jul-Aug;61(4):376-84. doi: 10.1002/bab.1187. Epub 2014 Mar 20.

Abstract

Bispecific antibody (BsAb) has been proved to be a very effective antitumor approach because of its distinctive advantages of immune-mediated cytotoxicity. To enhance the ability to recruit and activate T lymphocytes for tumor-specific killing, we constructed and prepared a recombinant human single-chain Fv bispecific antibody (BsAb), named VEGFR1/CD3 BsAb, targeting VEGFR1 and CD3. The VEGFR1/CD3 BsAb was expressed in CHO-K1 cells and purified by Ni-NTA affinity chromatography. The CD3 and VEGFR1-binding activity of VEGFR1/CD3 BsAb was confirmed by flow cytometry. T lymphocyte activation and proliferation induced by VEGFR1/CD3 BsAb were also demonstrated in vitro. Notably, our VEGFR1/CD3 BsAb presented a powerful and specific killing effect against VEGFR1-positive human breast cancer cell MDA-MB-231 and MDA-MB-435 through activating T lymphocyte at very low concentrations, indicating that it will be a valuable antibody drug for treatment of VEGFR1-positive cancers in the future.

Keywords: T lymphocyte; VEGFR1/CD3 BsAb; antibody drug; immunotherapy; tumor.

MeSH terms

  • Animals
  • Antibodies, Bispecific / biosynthesis*
  • Antibodies, Bispecific / immunology*
  • Antibodies, Bispecific / therapeutic use
  • Breast Neoplasms / drug therapy*
  • Breast Neoplasms / immunology*
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / pathology
  • CD3 Complex / immunology*
  • CHO Cells
  • Cell Line, Tumor
  • Cell Proliferation
  • Cricetulus
  • Drug Screening Assays, Antitumor
  • Humans
  • Vascular Endothelial Growth Factor Receptor-1 / immunology*

Substances

  • Antibodies, Bispecific
  • CD3 Complex
  • Vascular Endothelial Growth Factor Receptor-1