Cancer Immunology, Immunotherapy

, Volume 63, Issue 9, pp 877–888

Generation and characterization of a novel human IgG1 antibody against vascular endothelial growth factor receptor 2

  • Wei Xie
  • Daojuan Li
  • Juan Zhang
  • Zhike Li
  • Desmond Omane Acheampong
  • Yuan He
  • Youfu Wang
  • Zhiguo Chen
  • Min Wang
Original Article

DOI: 10.1007/s00262-014-1560-9

Cite this article as:
Xie, W., Li, D., Zhang, J. et al. Cancer Immunol Immunother (2014) 63: 877. doi:10.1007/s00262-014-1560-9

Abstract

VEGF and its receptors, especially VEGFR2 (KDR), are known to play a critical role in angiogenesis under both physiological and pathological conditions, including cancer and angiogenic retinopathies. This study was aimed at developing a fully human IgG1 antibody (mAb-04) constructed from a phage-derived scFv, targeting the VEGF/VEGFR2 pathway. Firstly, an innovative transfection system, containing two recombinant expression vectors (pMH3 and pCApuro), were introduced into CHO-s cells and clones with higher yield selected accordingly. After an optimal fermentation condition was determined, fed-batch fermentation was performed in 5-L bioreactor with a final yield up to 60 mg/L. Further, cell proliferation, wound healing, transwell invasion, tube formation and chick embryo chorioallantoic membrane assays showed significant anti-angiogenic activity of mAb-04 in vitro and in vivo. In addition, the results of Western blotting indicated the ability of mAb-04 to inhibit VEGF-induced VEGFR2 signaling pathway. Finally, ADCC assay demonstrated that mAb-04 is capable of mediating tumor cell killing in presence of effector cells. This study has therefore proved that the full-length antibody targeting human VEGFR2 has potential clinical applications in the treatment of cancer and other diseases where pathological angiogenesis is involved.

Keywords

VEGFR2 (KDR)Human monoclonal antibodyCHO cell lineAnti-angiogenesisADCC

Abbreviations

ADCC

Antibody-dependent cellular cytotoxicity

bFGF

Basic fibroblast growth factor

CAM

Chick embryo chorioallantoic membrane

ECGS

Endothelial cell growth supplement

ECM

Endothelial culture medium

FBS

Fetal bovine serum

HAMA

Human anti-mouse antibody

KDR

Kinase insert domain-containing receptor

KDR3

The extracellular domain 3 of human VEGFR2

LDH

Lactate dehydragenase

scFv

Single-chain antibody fragment

VEGF

Vascular endothelial growth factor

VEGFR2

Vascular endothelial growth factor receptor 2

Supplementary material

262_2014_1560_MOESM1_ESM.pdf (477 kb)
Supplementary material 1 (PDF 477 kb)

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Wei Xie
    • 1
  • Daojuan Li
    • 1
  • Juan Zhang
    • 1
  • Zhike Li
    • 1
  • Desmond Omane Acheampong
    • 1
  • Yuan He
    • 1
  • Youfu Wang
    • 1
  • Zhiguo Chen
    • 1
  • Min Wang
    • 1
  1. 1.State Key Laboratory of Natural Medicines, School of Life Science and TechnologyChina Pharmaceutical UniversityNanjingPeople’s Republic of China