Abstract:
VEGF and its receptors, especially VEGFR2
(KDR), are known to play a critical role in angiogen-
esis 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, target-
ing the VEGF/VEGFR2 pathway. Firstly, an innovative
transfection system, containing two recombinant expres-
sion vectors (pMH3 and pCApuro), were introduced into
CHO-s cells and clones with higher yield selected accord-
ingly. After an optimal fermentation condition was deter-
mined, 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 for-
mation 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 blot-
ting 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.
