Quantitative mass spectrometric immunoassay of insulin like growth factor 1

Randall W. Nelson, Dobrin Nedelkov, Kemmons A. Tubbs, Urban A. Kiernan

Research output: Contribution to journalArticlepeer-review

70 Scopus citations

Abstract

Reported in this work are the development of mass spectrometric immunoassay (MSIA) devices and methods for the qualitative analysis of IGF-1 and -2, and the rigorous quantification of IGF-1 from human plasma. A method involving addition of SDS in moderate concentration to unfractionated plasma for disrupting IGF/IGFBP complexes was initially developed. The method is suitable for the direct extraction of the IGFs and subsequent mass spectrometric analysis. Rat plasma, containing IGF-1 that is mass shifted from human IGF-1, was used as an internal reference standard (IRS) for the quantification of IGF-1 directly from human plasma. A standard curve with linear dynamic range of at least 2 orders of magnitude was constructed from serially diluted IGF-1 standards containing equal amounts of rat plasma. Using the standard curve, IGF-1 levels in plasma samples from eight individuals were determined. The limit of detection for the IGF-1 MSIA was also evaluated and established to be ∼15 pM. The assay is rapid and can be performed in parallel via high-throughput robotics processing. Furthermore, the mass spectrometry aspect of the developed IGF-1 immunoassay offers a new dimension in the ongoing study of IGF-1 and related diseases.

Original languageEnglish (US)
Pages (from-to)851-855
Number of pages5
JournalJournal of Proteome Research
Volume3
Issue number4
DOIs
StatePublished - Jul 1 2004

Keywords

  • IGF
  • Immunoassay
  • Mass spectrometry
  • Quantification

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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