Measurement of insulin-like growth factor-II in physiological fluids and tissues. I. An improved extraction procedure and radioimmunoassay for human and rat fluids

Ronald R. Bowsher, Wei Hua Lee, John M. Apathy, Peter J. O'Brien, Albert L. Ferguson, David P. Henry

Research output: Contribution to journalArticle

125 Citations (Scopus)

Abstract

The measurement of serum insulin-like growth factors (IGFs) in serum is complicated by the presence of high affinity IGF-binding proteins. The accurate measurement of IGFs by radioligand binding assays requires that the interference from binding proteins be eliminated. Acid-gel chromatography, the standard method for removing binding proteins, is laborious and time consuming. Alternative methods for extracting serum IGFs include the use of HCl-ethanol treatment and reverse phase minicolumns. However, these methods are unsuitable for use with serum for some species, such as rat and sheep, due to incomplete removal of binding proteins. We developed a fast protein liquid chromatography size-exclusion chromatographic method for characterizing the presence of IGF-binding proteins in physiological fluids and used this method to systematically investigate different combinations of acids and organic solvents as potential extraction methods for IGFs. We developed and validated an improved extraction procedure that uses formic acid, Tween-20, and acetone. The new extraction method was used in conjunction with purified biosynthetic human IGF-II and a commercially available anti-IGF-II monoclonal antibody in the development of an improved RIA for IGF-II. The new RIA is sensitive (5.0 pg/tube), specific (IGF-I cross-reactivity,

Original languageEnglish (US)
Pages (from-to)805-814
Number of pages10
JournalEndocrinology
Volume128
Issue number2
StatePublished - Feb 1991
Externally publishedYes

Fingerprint

Insulin-Like Growth Factor II
Radioimmunoassay
Somatomedins
Insulin-Like Growth Factor Binding Proteins
Carrier Proteins
formic acid
Serum
Radioligand Assay
Acids
Polysorbates
Acetone
Insulin-Like Growth Factor I
Liquid Chromatography
Gel Chromatography
Sheep
Ethanol
Monoclonal Antibodies

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Bowsher, R. R., Lee, W. H., Apathy, J. M., O'Brien, P. J., Ferguson, A. L., & Henry, D. P. (1991). Measurement of insulin-like growth factor-II in physiological fluids and tissues. I. An improved extraction procedure and radioimmunoassay for human and rat fluids. Endocrinology, 128(2), 805-814.

Measurement of insulin-like growth factor-II in physiological fluids and tissues. I. An improved extraction procedure and radioimmunoassay for human and rat fluids. / Bowsher, Ronald R.; Lee, Wei Hua; Apathy, John M.; O'Brien, Peter J.; Ferguson, Albert L.; Henry, David P.

In: Endocrinology, Vol. 128, No. 2, 02.1991, p. 805-814.

Research output: Contribution to journalArticle

Bowsher, Ronald R. ; Lee, Wei Hua ; Apathy, John M. ; O'Brien, Peter J. ; Ferguson, Albert L. ; Henry, David P. / Measurement of insulin-like growth factor-II in physiological fluids and tissues. I. An improved extraction procedure and radioimmunoassay for human and rat fluids. In: Endocrinology. 1991 ; Vol. 128, No. 2. pp. 805-814.
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