Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A next generation peripherally restricted Cavα2δ-1 ligand with inhibitory action on Cav2.2 channels and utility in neuropathic pain.

doi: 10.1016/j.biopha.2024.116472

Figure Lengend Snippet: Fig. 3. Specific SPR binding of the recombinant Cavα2δ-1 fragment comprising the VGGC_α2 domain to NVA1309, covalently coupled to the surface of a Biacore CM5 optical sensor chip (surface density 20 RU) (a). Binding sensorgram obtained with CUSABIO fragment; specificity is confirmed by lack of binding of non related proteins (human IgG and rec. human leukemia inhibiting factor (LIF), respectively). The binding sensorgram was analysed by mathematical curve fitting applying a Langmuir 1:1 interaction algorithm (BiaEvaluation 4.1 software). The fitted curve is shown in (b). Binding of human recombinant full length wild type and mutated α2δ-1 to NVA1309, covalently immobilised on the surface of a Biacore CM5 optical sensor chip: Each Cavα2δ-1 recombinant protein was run as analyte at three concentrations across the NVA1309 sensor chip surface. (NVA1309 surface density = 204RU). Kinetic binding constants and affinities (ka, kd; KD) were determined from double referenced sensorgrams by mathematical sensorgram fitting, with fitted curves shown in c-f.

Article Snippet: After covalent immobilisation of the fragments to flow cell surfaces FC2 and FC3 of a Biacore CM5 optical sensor chip, NVA1309 was passed as binding analyte at increasing concentrations over the ligand surfaces and a blank amine activated background reference surface FC1.

Techniques: Binding Assay, Recombinant, Software