Isotope dilution-mass spectrometric quantification of specific proteins: Model application with apolipoprotein A-I

John R. Barr, Vincent L. Maggio, Donald G. Patterson, Gerald R. Cooper, L. Omar Henderson, Wayman E. Turner, S. Jay Smith, W. Harry Hannon, Larry L. Needham, Eric J. Sampson

Research output: Contribution to journalArticle

312 Citations (Scopus)

Abstract

An enzymatic hydrolysis isotope dilution-mass spectrometric method was developed for reference quantification of specific proteins. The analytical procedure involved measuring a reproducibly hydrolyzed peptide (serving as the primary standard) unique to a specific protein. This new mass spectrometric method was evaluated by assessing the concentration of apolipoprotein (apo) A-I in the European Community Bureau of Reference (BCR) lyophilized Certified Reference Material (CRM 393). We used the method to make 96 measurements (4 replicate analyses of 4 enzymatic digests of 6 vials of BCR-CRM 393), which gave an average total protein mass of 1.048 mg (±1.0% at 99% confidence limits). The total overall analytical CV was 3.95%. The results of this evaluation of our model approach to determine the concentration of a specific protein in a purified preparation demonstrated that our new mass spectrometric method can be used to measure apolipoproteins and other specific proteins without the use of epitopic immunoassay methods.

Original languageEnglish (US)
Pages (from-to)1676-1682
Number of pages7
JournalClinical Chemistry
Volume42
Issue number10
StatePublished - Nov 2 1996

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Apolipoprotein A-I
Isotopes
Dilution
Proteins
Enzymatic hydrolysis
Apolipoproteins
European Union
Immunoassay
Hydrolysis
Peptides

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Barr, J. R., Maggio, V. L., Patterson, D. G., Cooper, G. R., Henderson, L. O., Turner, W. E., ... Sampson, E. J. (1996). Isotope dilution-mass spectrometric quantification of specific proteins: Model application with apolipoprotein A-I. Clinical Chemistry, 42(10), 1676-1682.
Barr, John R. ; Maggio, Vincent L. ; Patterson, Donald G. ; Cooper, Gerald R. ; Henderson, L. Omar ; Turner, Wayman E. ; Smith, S. Jay ; Hannon, W. Harry ; Needham, Larry L. ; Sampson, Eric J. / Isotope dilution-mass spectrometric quantification of specific proteins : Model application with apolipoprotein A-I. In: Clinical Chemistry. 1996 ; Vol. 42, No. 10. pp. 1676-1682.
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Barr, JR, Maggio, VL, Patterson, DG, Cooper, GR, Henderson, LO, Turner, WE, Smith, SJ, Hannon, WH, Needham, LL & Sampson, EJ 1996, 'Isotope dilution-mass spectrometric quantification of specific proteins: Model application with apolipoprotein A-I', Clinical Chemistry, vol. 42, no. 10, pp. 1676-1682.

Isotope dilution-mass spectrometric quantification of specific proteins : Model application with apolipoprotein A-I. / Barr, John R.; Maggio, Vincent L.; Patterson, Donald G.; Cooper, Gerald R.; Henderson, L. Omar; Turner, Wayman E.; Smith, S. Jay; Hannon, W. Harry; Needham, Larry L.; Sampson, Eric J.

In: Clinical Chemistry, Vol. 42, No. 10, 02.11.1996, p. 1676-1682.

Research output: Contribution to journalArticle

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AU - Needham, Larry L.

AU - Sampson, Eric J.

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Barr JR, Maggio VL, Patterson DG, Cooper GR, Henderson LO, Turner WE et al. Isotope dilution-mass spectrometric quantification of specific proteins: Model application with apolipoprotein A-I. Clinical Chemistry. 1996 Nov 2;42(10):1676-1682.