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Acid-Labile Subunit of Human Insulin-Like Growth Factor-Binding Protein Complex: Measurement, Molecular, and Clinical Evaluation

Diagnostic Systems Laboratories (Canada), Inc. (M.J.K., A.D.), and the Department of Clinical Biochemistry, University of Toronto (M.J.K.), Toronto, Ontario, Canada; and Diagnostic Systems Laboratories, Inc. (J.M., R.G.K., A.K.), Webster, Texas 77598

Address all correspondence and requests for reprints to: M. J. Khosravi, Ph.D., Diagnostic Systems Laboratories (Canada), Inc., Mount Sinai Hospital, Room 653, 600 University Avenue, Toronto, Ontario, Canada M5G 1X5.

Although the acid-labile subunit (ALS) of the 150-kDa insulin-like growth factor (IGF)-binding protein (IGFBP) complex was described over a decade ago, details of ALS physiology have remained largely uncertain. We evaluated antibodies to synthetic human ALS and constructed a noncompetitive ALS enzyme-linked immunosorbent assay. Whereas uncomplexed ALS is directly measured, determination of total levels required sample pretreatment with SDS, which was found to optimally dissociate complexed ALS and significantly enhance ALS immunoreactivity. ALS in random adult sera was approximately 50% uncomplexed, and samples devoid of complexed ALS by immunoaffinity separation contained about 54% of the total levels. Serum ALS fractionated by gel filtration high performance liquid chromatography eluted in a single peak at approximately 150 kDa with IGF-I and IGFBP-3, but appeared at about 400–500 kDa after sample acidification and fractionation under acidic condition. The unexpected shift in ALS immunoreactivity remained unchanged when acid-neutralized or SDS-treated samples were fractionated under neutral pH and was reproducible when the 150-kDa complex was isolated, treated with acid or SDS, and rechromatographed. ALS in adult sera more tightly correlated with IGFBP-3 than IGF-I or IGF-II. The total levels (mean ± SD) were 16.7 ± 3.7 mg/L in 22 normal subjects, 28.3 ± 8.1 mg/L in 20 acromegalic patients, and 9.5 ± 3.8 in 32 GH-deficient adults. Little or no ALS was detectable in amniotic fluid, cerebrospinal fluid, seminal plasma, or milk, whereas high levels were present in synovial fluid. The development of ALS enzyme-linked immunosorbent assay should greatly facilitate further investigations of this unique glycoprotein.

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