Homogeneous insulin and C-Peptide sensors for rapid assessment of insulin and C-peptide secretion by the islets. Diabetes 2010 Oct;59(10):2360-5
Date
07/14/2010Pubmed ID
20622164Pubmed Central ID
PMC3279555DOI
10.2337/db10-0088Scopus ID
2-s2.0-77957580825 (requires institutional sign-in at Scopus site) 38 CitationsAbstract
OBJECTIVE: 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.
RESEARCH DESIGN AND METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
Author List
Heyduk E, Moxley MM, Salvatori A, Corbett JA, Heyduk TAuthor
John A. Corbett PhD Chair, Professor in the Biochemistry department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AntibodiesBase Sequence
Biosensing Techniques
C-Peptide
Enzyme-Linked Immunosorbent Assay
Humans
Insulin
Islets of Langerhans
Limit of Detection
Oligodeoxyribonucleotides
Proinsulin
Radioimmunoassay
