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 VEGFinduced
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.
