Journal: Pharmaceutics
Article Title: Brain Delivery of IGF1R5, a Single-Domain Antibody Targeting Insulin-like Growth Factor-1 Receptor
doi: 10.3390/pharmaceutics14071452
Figure Lengend Snippet: In vitro and in vivo BBB transport of IGF1R5 in the V H H format or in hFc or mFc fusions. ( A ) Transport of IGF1R5 in the V H H format, and humanized (H2) and hFc fused variants of IGF1R5 across a rat (SV-ARBEC) and human (iBEC) in vitro BBB model. Antibodies were applied in the upper compartment of the BBB insert and then quantified over time in the bottom chamber using SRM to determine Papp values. Papp values (cm/min) of antibodies are shown as means ± sd derived from 6 separate transwell inserts. * p < 0.05 vs. respective negative control. ( B ) Transport of mFc-IGF1R5 and IGF1R5-mFc across a rat BBB model in vitro. Antibodies were paired with corresponding controls in the upper compartment of the BBB insert and then quantified over time in the bottom chamber using SRM to determine the Papp values. Papp values (cm/min) of antibodies are shown as means ± sd derived from 6 separate transwell inserts. * p < 0.05 vs. respective negative control. ( C ) Concentrations of hFc-IGF1R5 or A20.1-mFc in serum, CSF or capillary-depleted brain of rats at 24 h following a bolus i.v. injection of 15 mg/kg of each antibody. The concentrations were measured using SRM analysis in at least 3 animals and the bars represent mean and SD. * p < 0.01 vs. A20.1-mFc. ( D – G ) Immunofluorescence staining of the rat frontal cortex 48 h after intravenous injection of 15 mg/kg of IGF1R5-mFc ( D – F ) or an equimolar dose of A20.1-mFc ( G , H ). Brain vessels were detected using RCA-1 (green); neurons were detected using antibodies against NeuN (blue); IGF1R5-mFc (red). Images were observed at 10x objective ( D ) and 60x ( E – H ). Scale bars, 100 µm and 10 µm.
Article Snippet: The BBB permeability of IGF1R5 in the V H H format, the humanized H2 variant of IGF1R5 and the hFc-IGF1R5H2, along with their respective negative controls, were assessed in rat (SV-ARBEC) and human (iBEC) BBB models in vitro.
Techniques: In Vitro, In Vivo, Derivative Assay, Negative Control, Injection, Immunofluorescence, Staining
