Homogeneous Insulin and C-Peptide Sensors for Rapid Assessment of Insulin and C-Peptide Secretion by the Islets
Open Access
- 9 July 2010
- journal article
- Published by American Diabetes Association in Diabetes
- Vol. 59 (10), 2360-2365
- https://doi.org/10.2337/db10-0088
Abstract
Glucose-stimulated islet insulin or C-peptide secretion experiments are a fundamental tool for studying and assessing islet function. The goal of this work was to develop Ab-based fluorescent homogenous sensors that would allow rapid, inexpensive, near-instantaneous determinations of insulin and C-peptide levels in biological samples. Our approach was to use molecular pincer design (Heyduk et al., Anal Chem 2008;80:5152–5159) in which a pair of antibodies recognizing nonoverlapping epitopes of the target are modified with short fluorochrome-labeled complementary oligonucleotides and are used to generate a fluorescence energy transfer (FRET) signal in the presence of insulin or C-peptide. The sensors were capable of detecting insulin and C-peptide with high specificity and with picomolar concentration detection limits in times as short as 20 min. Insulin and C-peptide levels determined with the FRET sensors showed outstanding correlation with determinations performed under the same conditions with enzyme-linked immunosorbent assay. Most importantly, the sensors were capable of rapid and accurate determinations of insulin and C-peptide secreted from human or rodent islets, verifying their applicability for rapid assessment of islet function. The homogeneous, rapid, and uncomplicated nature of insulin and C-peptide FRET sensors allows rapid assessment of β-cell function and could enable point-of-care determinations of insulin and C-peptide.Keywords
This publication has 13 references indexed in Scilit:
- β-cell transplantation for diabetes therapyThe Lancet, 2008
- Safety of aprotinin (again)The Lancet, 2008
- Molecular Pincers: Antibody-Based Homogeneous Protein SensorsAnalytical Chemistry, 2008
- International Trial of the Edmonton Protocol for Islet TransplantationThe New England Journal of Medicine, 2006
- Edmonton's islet success has indeed been replicated elsewhereThe Lancet, 2003
- Isolation of Islets of Langerhans from Rodent PancreasMethods in molecular medicine, 2002
- Human Islet Transplantation: UpdateWorld Journal of Surgery, 2001
- Islet Transplantation in Seven Patients with Type 1 Diabetes Mellitus Using a Glucocorticoid-Free Immunosuppressive RegimenThe New England Journal of Medicine, 2000
- Thiol-Reactive, Luminescent Europium Chelates: Luminescence Probes for Resonance Energy Transfer Distance Measurements in BiomoleculesAnalytical Biochemistry, 1997
- [13] Fluorescence resonance energy transferMethods in Enzymology, 1995