Over-expression of vascular endothelial growth factor (vegf) leads to the proliferation and neovasculature of cancer cells. targeting of vegf signaling is effective way to reduce or inhibit tumor proliferation and angiogenesis. appropriate pharmaceutical properties of came single domain antibodies (known as nanobody) like: single domain entity and ease to manipulation make them potential for drug development studies. here we constructed a bi-specific format (targeting two distinct epitopes on vegf) of previously developed nanobody to efficiently target tumor cells proliferation.
Bi-specific nanobody designed and cloned into wk6 e.coli cells. expression was induced by 0.5mm iptg and monitored by sds-page and western blot analysis. functional assay of the nanobody was performed on human endothelial cells by mtt and tube formation assay.
Results showed that the bi-specific nanobody significantly inhibited proliferation and tube formation of endothelial cells in compare to mono-specific nanobody through blockade of vegf signaling.
Taken together, the bi-specific nanobody has potential to introduce as novel therapeutic in treatment of angiogenesis-dependent disease like caner.