Production and characterization of a high-affinity nanobody against human endoglin.
Abstract Antibodies or antibody fragments are almost exclusively applied in human therapy and diagnosis. The high affinity and specificity of antibodies makes them suitable for these applications. Nanobody, the variable domain of Camelidae heavy chain antibodies, have superior properties compared with conventional antibodies in that they are small, non-immunogenic, very stable, highly soluble, and easy to produce in large quantities. In the present study, we report the isolation and characterization of a high-affinity binder against human endoglin retrieved from camels' nanobody gene library. Endoglin (CD105), an accessory protein of the transforming growth factor beta receptor complex, has become an attractive molecule for the targeting of the tumor vasculature. Upregulation of endoglin on proliferating endothelial cells is associated with tumor neovascularization. Here, we generated two nanobody gene libraries displayed on phage particles. Some single-domain antibody fragments have been isolated that specifically recognize the recombinant extracellular domain of human endoglin. The other selected anti-endoglin nanobody (AR1-86) showed strong binding to human endoglin expressing endothelial cells (HUVECs), while no binding was observed with the endoglin-negative cell line (HEK293). This high-affinity single-domain antibody could be a good candidate for the generation of vascular or tumor targeting agents in cancer therapy.
Department of Clinical Biochemistry, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Davoud
D
Ahmadvand
Mohammad J
MJ
Rasaee
Fatemeh
F
Rahbarizadeh
Mohammad
M
Mohammadi
D000818:Animals; D000918:Antibody Specificity; D015703:Antigens, CD / Q000276:immunology*; D002162:Camelus; D002478:Cells, Cultured; D000071063:Endoglin; D042783:Endothelial Cells / Q000276:immunology; D004730:Endothelium, Vascular / Q000276:immunology; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D019151:Peptide Library; D011956:Receptors, Cell Surface / Q000276:immunology*
D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011956:Receptors, Cell Surface