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: SPR-derived kinetics and equilibrium dissociation constants for V H H-IGF1R interactions. 1 Determined at pH 7.4, 25 °C; 2 determined at pH 5.6, 37 °C.

Article Snippet: Ectodomains of human IGF1R (R&D Systems, Cat#391-GR-050), rhesus IGF1R (NRC Montreal, aa31-932), rat IGF1R (NRC Montreal, aa31-936) and mouse IGF1R (R&D Systems, Cat#6630-GR/CF-025) were amine coupled on CM5 sensor chips (Cytiva) at 10 μg/mL in 10 mM acetate pH 4.0 using an amine-coupling kit (Cytiva), resulting in approximately 1500–2000 response units (RUs) of each IGF1R ectodomain immobilized.

Techniques:

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: High-affinity binding of IGF1R5 V H Hs to IGF1R ectodomains. ( A ) SPR sensorgrams demonstrating wild-type IGF1R5 and humanized IGF1R5-H2 V H Hs binding to surface-immobilized human, rhesus, mouse and rat IGF1R (pH 7.4, 25 °C). V H H concentrations in flow ranged from 0.25 to 10 nM (IGF1R5) and from 1 to 25 nM (IGF1R5-H2). Kinetics and affinities were determined using multi-cycle kinetics (human, mouse, rat IGF1R) or single-cycle kinetics (rhesus IGF1R) analyses. ( B ) Sensorgrams demonstrating the binding of V H Hs to human and mouse IGF1R at acidic pH (pH 5.6, 37 °C). V H H concentrations in flow ranged from 0.25 to 10 nM (IGF1R5) and from 1 to 50 nM (IGF1R5-H2). Black lines: raw data; red lines: 1:1 binding model fitting.

Article Snippet: Ectodomains of human IGF1R (R&D Systems, Cat#391-GR-050), rhesus IGF1R (NRC Montreal, aa31-932), rat IGF1R (NRC Montreal, aa31-936) and mouse IGF1R (R&D Systems, Cat#6630-GR/CF-025) were amine coupled on CM5 sensor chips (Cytiva) at 10 μg/mL in 10 mM acetate pH 4.0 using an amine-coupling kit (Cytiva), resulting in approximately 1500–2000 response units (RUs) of each IGF1R ectodomain immobilized.

Techniques: Binding Assay