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Updated: Aug 6, 2026
Homogeneous Time-resolved Förster Resonance Energy Transfer-based Assay for Detection of Insulin Secretion
Published on: May 10, 2018
Analysis of solvent-mediated conformational changes of insulin by radioimmunoassay (RIA) techniques
S P Musial1, M P Duran, R V Smith
1Drug Dynamics Institute and Division of Medicinal Chemistry, College of Pharmacy, University of Texas at Austin, Austin, TX 78712, USA.
Stationary phase radioimmunoassay (RIA) (i.e. antibodies bound to polystyrene test tubes) techniques are used as an analytical probe of secondary and tertiary structural changes of radiolabelled (125)I porcine insulin. The effects of temperature, buffer composition, pH and ionic strength and solvents on insulin binding are studied. Optimum insulin-antibody binding occurred at 22 degrees C, pH 6 and a buffer strength of 0.1 M or less. Results of experiments with three pH 6 buffers (0.005 M phosphate, 0.1 M acetate and 0.1 M Tris) showed no statistical difference in binding properties. For all solvents tested, increasing the solvent concentration decreased the amount of insulin binding. Comparison of the various solvents tested indicated that ethylene glycol and methanol are the least denaturant whilst 1-propanol and acetonitrile are among the most denaturant.
Stationary phase radioimmunoassay (RIA) (i.e. antibodies bound to polystyrene test tubes) techniques are used as an analytical probe of secondary and tertiary structural changes of radiolabelled (125)I porcine insulin. The effects of temperature, buffer composition, pH and ionic strength and solvents on insulin binding are studied. Optimum insulin-antibody binding occurred at 22 degrees C, pH 6 and a buffer strength of 0.1 M or less. Results of experiments with three pH 6 buffers (0.005 M phosphate, 0.1 M acetate and 0.1 M Tris) showed no statistical difference in binding properties. For all solvents tested, increasing the solvent concentration decreased the amount of insulin binding. Comparison of the various solvents tested indicated that ethylene glycol and methanol are the least denaturant whilst 1-propanol and acetonitrile are among the most denaturant.
