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This version published online on April 19, 2005
Journal of Clinical Endocrinology & Metabolism, doi:10.1210/jc.2004-2090
A more recent version of this article appeared on July 1, 2005
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Submitted on October 22, 2004
Accepted on April 11, 2005

A RAPID METHOD FOR ANALYZING SERUM PRO-INSULIN-LIKE GROWTH FACTOR-II (PRO-IGF-II) IN PATIENTS WITH NON-ISLET CELL TUMOUR HYPOGLYCEMIA

Farideh Miraki-Moud, Ashley B Grossman, Michael Besser, John P Monson, and Cecilia Camacho-Hübner*

Department of Endocrinology, William Harvey Research Institute, Queen Mary University of London, U.K.

* To whom correspondence should be addressed. E-mail: c.camacho-hubner{at}qmul.ac.uk.

Context: Non-islet cell tumor hypoglycemia (NICTH) results from the hypersecretion of pro-IGF-II by a large, usually, mesenchymal tumor. Detection of pro-IGF-II in serum is a potential tumor marker in these patients.

Objective: To validate a rapid and reliable method for determining serum pro-IGF-II. Patients Serum samples from 16 patients with NICTH were studied.

Main Outcome Measures: Serum concentration of pro-IGF-II determined by immunoblot analysis of pro-IGF-II and mature IGF-II after 16.5% tricine-SDS-PAGE (tricine SDS-PAGE) which was compared with pro-IGF-II measured by standard RIA after size-exclusion acid chromatography.

Results: The analyses of patients sera by size-exclusion acid chromatography showed that 68 ± 19% of IGF-II were present in the pro-IGF-II form, whereas only 18 ± 4% corresponded to pro-IGF-II in controls. Scanning densitometry of immunoblots showed 67 ± 16% in the bands corresponding to pro-IGF-II in patients sera compared with 27 ± 9% in controls. The detection sensitivity of tricine SDS-PAGE method was the same as for size-exclusion chromatography, but the tricine SDS-PAGE method is quicker and requires smaller amounts of serum.

Conclusion: Tricine SDS-PAGE followed by IGF-II immunoblot analysis provides a rapid, reproducible and sensitive method for the separation of serum pro-IGF-II from mature IGF-II, and is a useful laboratory evaluation of patients with a clinical diagnosis of NICTH.
Key words: Non-islet cell tumour hypoglycemia • pro-insulin-like growth factor-II (pro-IGF-II) • tricine-sodium dodecyl sulfate polyacrylamide gel electrophoresis (tricine SDS-PAGE)




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