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Differential removal of insulin-like growth factor binding proteins in rat serum by solvent extraction procedures

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Abstract

Solvent extraction of serum and other biological fluids at an acidic pH is a convenient method to remove the insulin-like growth factor binding proteins (IGFBPs); however, an incomplete removal of IGFBPs can occur and this can potentially interfere with the radioimmunoassay of insulin-like growth factors (IGFs). This study compared the removal of IGFBPs from normal adult rat serum and 5-day old neonatal rat serum by acid-gel filtration, and three solvent extraction methods, i.e., acid-ethanol (AE), acid-cryo-ethanol (ACE) and formic acid-acetone (FAA) treatments by western ligand blotting and slot-blotting analysis. In adult rat serum all three extraction methods removed nearly 75% of total IGFBPs present. For the neonatal serum, AE and FAA were very inefficient in eliminating the IGFBPs, while ACE was somewhat better, as it removed nearly 30% of IGFBPs. Ligand blots of extracted samples showed that IGFBPs of lower size range, 24 to 32 kDa (IGFBP-4, IGFBPs-1 and-2), were resistant to solvent extraction. Acid-gel filtration, in contrast, eliminated >95% of IGF-binding components in both sera. Determination of IGF-I concentrations in samples after gel filtration and extraction methods revealed lower IGF-I values in neonatal serum in acid extracted samples. These data caution against using solvent extractions for IGFBP removal in fetal/neonatal serum.

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Authors and Affiliations

  1. Endocrine-Metabolic Division, Veterans Affairs Medical Center and The Chicago Medical School, 60064, North Chicago, Illinois, USA

    K. Srivenugopal, S. P. Singh M.D., X. -H. Yuan, S. Ehmann & A. K. Snyder

  2. VA Medical Center, 3001 Green Bay Road, 60064, North Chicago, IL, USA

    S. P. Singh M.D.

Authors
  1. K. Srivenugopal
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  2. S. P. Singh M.D.
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  3. X. -H. Yuan
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  4. S. Ehmann
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  5. A. K. Snyder
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Srivenugopal, K., Singh, S.P., Yuan, X.H. et al. Differential removal of insulin-like growth factor binding proteins in rat serum by solvent extraction procedures. Experientia 50, 451–455 (1994). https://doi.org/10.1007/BF01920745

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  • DOI: https://doi.org/10.1007/BF01920745

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