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Clinical Chemistry 21: 873-879, 1975;
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Clinical Chemistry, Vol 21, 873-879, Copyright © 1975 by the American Association for Clinical Chemistry

Quantitation of Free, Total, and Antibody-bound Insulin in Insulin-Treated Diabetics

William D. Gennaro 1 and John D. Van Norman 1

1 Department of Chemistry, Youngstown State University, Youngstown, Ohio 44555.

We describe a simplified method for measuring free, total, and antibody-bound insulin in insulin-treated patients in whom antibodies to insulin are present. The free, active insulin is extracted from the serum with a polyethylene glycol solution. Total insulin is extracted from the serum with a polyethylene glycol solution after dissociation of the antibody—antigen complex with dilute HCl. Aliquots of the extracts are used in the radioimmunoassay system. The figure for antibody-bound insulin is the difference between the total and free insulin values and reflects the concentration of insulin antibodies present. A commercially available ("Phadebas") radioimmunoassay for immunoreactive insulin was used to quantitate the insulin present in the two extracts. Recovery of added insulin averaged 85% for the free insulin and 87% for the total insulin.


Key Words: radioimmunoassay • "kit" method • reference values • non-effect of tolbutamide • analytical problems of insulin resistance • polyethylene glycol

Submitted on December 15, 1974
Accepted on March 24, 1975




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Copyright © 1975 by the American Association for Clinical Chemistry.