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1
Diagnostic Systems Laboratories (Canada) Inc., Toronto, ON, Canada.
2
Department of Clinical Biochemistry, University of
Toronto, Toronto, ON, Canada.
3
Diagnostic Systems Laboratories, Webster, TX.
a Address correspondence to this author at: Diagnostic Systems Laboratories (Canada) Inc., Mount Sinai Hospital, Room 653, 600 University Ave., Toronto, ON, Canada M5G 1X5. Fax 416-586-8479.
Accurate measurement of insulin-like growth factor (IGF) binding protein-1 (IGFBP-1) is important for precise definition of its physiological roles and potential diagnostic values. Because altered phosphorylation results in altered IGFBP-1 immunoreactivity, current assays may significantly underestimate or fail to detect physiological changes in the IGFBP-1 concentrations. We developed three ELISAs (ELISA 1–3) using a common capture but three different detection antibodies. IGFBP-1 in serum, synovial fluid (SF), cerebrospinal fluid (CSF), and amniotic fluid (AF) were measured before and after treatment with alkaline phosphatase (ALP). Among the methods, only ELISA-1 was unaffected by IGFBP-1 phosphorylation and generated identical results before and after ALP treatment. The serum and SF values by ELISA-2 and -3 were lower by ~4- to 10-fold, but increased after ALP treatment to within 66–98% of those by ELISA-1. The medians in AF, and to a lesser extent in CSF, by all methods were similar and did not change significantly after dephosphorylation. ELISA-1 showed excellent correlation with ELISA-2, ELISA-3, and a commercial IGFBP-1 IRMA only after ALP-treated samples were analyzed by the comparative methods. ELISA-1 is highly specific for IGFBP-1 and demonstrated acceptable analytical performance characteristics.
Key Words: indexing terms: insulin-like growth factors • insulin-like growth factor binding proteins • phosphorylation • ELISA
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