Immunoradiometric assay of human proinsulin and partially processed proinsulin with use of monoclonal antibody and streptavidin-biotin labeling.
10.3346/jkms.1989.4.4.171
- Author:
Jin Q KIM
1
;
Han Ik CHO
;
Sang In KIM
;
Hong Kyu LEE
;
C N HALES
Author Information
1. Department of Clinical Pathology, Seoul National University College of Medicine, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
immunoradiometric assay;
monoclonal antibodies;
streptavidin-biotin;
proinsulin;
diabetes
- MeSH:
Adult;
Animals;
Antibodies, Monoclonal/*immunology;
Bacterial Proteins;
Biotin;
Cross Reactions;
Humans;
Immunoradiometric Assay;
Male;
Mice;
Proinsulin/*analysis/immunology;
Reproducibility of Results;
Streptavidin
- From:Journal of Korean Medical Science
1989;4(4):171-177
- CountryRepublic of Korea
- Language:English
-
Abstract:
The sensitive and specific immunoradiometric assay is described for human proinsulin and its intermediate peptides (65-66 split and 32-33 split proinsulin). We developed a monoclonal antibody-based two-site immunoradiometric assay with use of streptavidin-biotin labeling. The detection limits of the assays lie in the range of 0.5-2.0 pM. In the proinsulin assay proinsulin cross-reacted 66% with 65-66 split proinsulin but not with insulin or 32-33 split proinsulin. In the assay of 65-66 split proinsulin it does not cross-react with insulin, proinsulin or 32-33 split proinsulin. In the 32-33 split proinsulin assay it cross-reacted 84% with proinsulin and 60% with 65-66 split proinsulin. The precision (C.V.) of the assays was less than 15% over the various concentration. The mean concentrations of insulin, proinsulin, 65-66 split proinsulin and 32-33 proinsulin in eight young male subjects in the fasting state were (pM +/- S.E.M.) 20 +/- 3.6, 2.3 +/- 0.3, undetectable (less than 1.0) and 2.1 +/- 0.7 and at the maximum reached during an oral glucose tolerance test, 150 +/- 26, 9.9 +/- 1.4, 3.8 +/- 0.6 and 19.7 +/- 6.0 respectively.
