[en] We describe a sensitive two-site sandwich enzyme-linked immunosorbent assay for the measurement of intact human proinsulin in 100 microL of serum or plasma. The assay is based on the use of two monoclonal antibodies specific for epitopes at the C-peptide/insulin A chain junction and at the insulin B chain/C-peptide junction, respectively. Cross-reactivities with insulin, C-peptide, and the four proinsulin conversion intermediates were negligible. The detection limit in buffer was 0.2 pmol/L (3 standard deviations from zero). The working range was 0.2-100 pmol/L. The mean intra- and interassay coefficients of variation were 2.4% and 8.9%, respectively. The mean recovery of added proinsulin was 103%. Dilution curves of 40 serum samples are parallel to the proinsulin calibration curve. Proinsulin concentrations in 20 fasting healthy subjects were all above the limit of detection: median (range), 2.7 pmol/L (1.1-6.9 pmol/L). Six fasting non-insulin-dependent diabetes mellitus and five insulinoma patients had proinsulin concentrations significantly higher than healthy subjects: median (range), 7.7 pmol/L (3.2-18 pmol/L) and 153 pmol/L (98-320 pmol/L), respectively.
Disciplines :
Endocrinology, metabolism & nutrition
Author, co-author :
Houssa, P.
Dinesen, B.
Deberg, Michelle ; Université de Liège - ULiège > Unité de recherche sur l'os et le cartillage (U.R.O.C.)
Frank, B. H.
Van Schravendijk, C.
Sodoyez-Goffaux, F.
Sodoyez, J. C.
Language :
English
Title :
First Direct Assay for Intact Human Proinsulin
Publication date :
July 1998
Journal title :
Clinical Chemistry
ISSN :
0009-9147
eISSN :
1530-8561
Publisher :
American Association for Clinical Chemistry, Washington, United States - District of Columbia
scite shows how a scientific paper has been cited by providing the context of the citation, a classification describing whether it supports, mentions, or contrasts the cited claim, and a label indicating in which section the citation was made.
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