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KMID : 0191119890040040171
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Journal of Korean Medical Science
1989 Volume.4 No. 4 p.171 ~ p.177
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Immunoradiometric assay of human proinsulin and partially processed proinsulin with use ofmonoclonal antibody and streptavidin-biotin labeling
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Kim JQ
Cho HI/Kim SI/Lee HK/Hales CN
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Abstract
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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.
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