title: Development of a feline proinsulin immunoradiometric assay and a feline proinsulin enzyme-linked immunosorbent assay (ELISA): a novel application to examine beta cell function in cats.
abstract: The prevalence of feline diabetes mellitus has increased several-fold over the last three decades. In humans, progression from obesity to diabetes is marked by changes in the release of proinsulin. A specific proinsulin (FPI) assay has not been available to examine similar changes in cats. The goal of this study was to develop a proinsulin assay for the analysis of beta cell function in cats. Monoclonal antibodies were developed against recombinant FPI and used in a two-site sandwich immunoradiometric assay (IRMA) and enzyme-linked immunosorbent Assay (ELISA). The antibody pair had negligible cross-reactivity with bovine insulin and feline C-peptide. The working range was 11-667pmol/L for the IRMA and 11-1111pmol/L for the ELISA. An intravenous glucose tolerance test was performed in six long-term obese and six lean adult healthy cats and serum glucose, insulin, and FPI concentrations were determined. The proinsulin and insulin secretion pattern in response to glucose was significantly different between lean and obese cats but the pattern was similar within a group. Both groups had similar baseline proinsulin/insulin ratios; however, obese cats showed a significantly higher proinsulin/insulin ratio during the first 15min of the IVGTT and a much lower ratio during the last 30min suggesting a time-delayed adjustment to the increased insulin demand. In conclusion, we report the development and validation of an IRMA and an ELISA for FPI. This novel assay is useful to elucidate FPI secretion and can be used similar to a C-petide assay to evaluate residual beta cell function in cats.
authors.affiliation: Department of Physiology and Pharmacology, University of Georgia, College of Veterinary Medicine, Athens, GA, United States.
authors.forename: S
authors.initials: S
authors.lastname: Kley
authors.forename: Z
authors.initials: Z
authors.lastname: Caffall
authors.forename: E
authors.initials: E
authors.lastname: Tittle
authors.forename: D C
authors.initials: DC
authors.lastname: Ferguson
authors.forename: M
authors.initials: M
authors.lastname: Hoenig
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000032:analysis; D002371:Cat Diseases / Q000097:blood / Q000175:diagnosis*; D002415:Cats; D003920:Diabetes Mellitus / Q000175:diagnosis / Q000503:physiopathology / Q000662:veterinary*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D005260:Female; D015309:Glucose Clamp Technique; D005951:Glucose Tolerance Test / Q000662:veterinary; D015592:Immunoradiometric Assay / Q000662:veterinary*; D007328:Insulin / Q000097:blood / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000502:physiology*; D008297:Male; D009765:Obesity / Q000097:blood / Q000662:veterinary; D011384:Proinsulin / Q000097:blood* / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D001786:Blood Glucose; D007328:Insulin; D011384:Proinsulin