Intraocular Penetration of a vNAR: In Vivo and In Vitro VEGF165 Neutralization.
Variable new antigen receptor domain (vNAR) antibodies are novel, naturally occurring antibodies that can be isolated from naïve, immune or synthetic shark libraries. These molecules are very interesting to the biotechnology and pharmaceutical industries because of their unique characteristics related to size and tissue penetrability. There have been some approved anti-angiogenic therapies for ophthalmic conditions, not related to vNAR. This includes biologics and chimeric proteins that neutralize vascular endothelial growth factor (VEGF)165, which are injected intravitreal, causing discomfort and increasing the possibility of infection. In this paper, we present a vNAR antibody against human recombinant VEGF165 (rhVEGF165) that was isolated from an immunized Heterodontus francisci shark. A vNAR called V13, neutralizes VEGF165 cytokine starting at 75 μg/mL in an in vitro assay based on co-culture of normal human dermal fibroblasts (NHDFs) and green fluorescence protein (GFP)-labeled human umbilical vein endothelial cells (HUVECs) cells. In the oxygen-induced retinopathy model in C57BL/6:Hsd mice, we demonstrate an endothelial cell count decrease. Further, we demonstrate the intraocular penetration after topical administration of 0.1 μg/mL of vNAR V13 by its detection in aqueous humor in New Zealand rabbits with healthy eyes after 3 h of application. These findings demonstrate the potential of topical application of vNAR V13 as a possible new drug candidate for vascular eye diseases.
['Camacho-Villegas|Tanya A|TA|', 'Mata-González|María Teresa|MT|', 'García-Ubbelohd|Walter|W|', 'Núñez-García|Linda|L|', 'Elosua|Carolina|C|', 'Paniagua-Solis|Jorge F|JF|', 'Licea-Navarro|Alexei F|AF|0000-0003-4022-7405']
D000287:Administration, Topical D000818:Animals D001688:Biological Products D002478:Cells, Cultured D018920:Coculture Techniques D004195:Disease Models, Animal D005123:Eye D005347:Fibroblasts D061307:Human Umbilical Vein Endothelial Cells D006801:Humans D008297:Male D051379:Mice D008810:Mice, Inbred C57BL D009883:Ophthalmic Solutions D010100:Oxygen D011817:Rabbits D011994:Recombinant Proteins D012164:Retinal Diseases D012754:Sharks D061905:Single-Domain Antibodies D042461:Vascular Endothelial Growth Factor A
intraocular penetration vNAR VEGF165 Heterodontus fransisci age-related macular degeneration diabetic retinopathy horn shark single chain binding domain