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Proteomics and Protein Markers |
1 HyTest Ltd., Turku, Finland.
2 Department of Biochemistry, Moscow State University, Moscow, Russia.
3 Moscow Research Institute of Medical Ecology, Moscow, Russia.
4 67 City Hospital, Moscow, Russia.
5 Moscow State Medico-Stomatological University, Moscow, Russia.
6 Department of Laboratory Medicine and Pathology, Hennepin County Medical Center, University of Minnesota School of Medicine, Minneapolis, MN.
aAddress correspondence to this author at: HyTest Ltd., Intelligate, 6th Floor, Joukahaisenkatu 6, 20520 Turku, Finland. Fax 358-25120909, e-mail alexei.katrukha{at}hytest.fi.
Background: Peptides derived from brain natriuretic peptide (BNP) precursor (proBNP), BNP, and the N-terminal fragment of proBNP (NT-proBNP) are used as biomarkers of heart failure. It remains unclear which forms of these peptides circulate in blood and which forms are measured by assays for these natriuretic peptides.
Methods: To design assays for immunodetection of proBNP, NT-proBNP, and BNP, we used a panel of BNP- and NT-proBNP–specific monoclonal antibodies (MAbs). All MAbs were tested in 2-site combinations in time-resolved fluoroimmunoassays with recombinant or synthetic antigens and plasma from heart failure (HF) patients. ProBNP and related molecules were assayed in HF plasma samples and plasma extracts by means of gel filtration fast protein liquid chromatography (FPLC) before and after protein fractionation on Sep-Pak C18 cartridges.
Results: The limits of detection for BNP, proBNP, and NT-proBNP assays were 0.4, 3, and 10 ng/L, respectively. Gel filtration-FPLC studies revealed 1 peak of NT-proBNP (
25 kDa), 1 peak of proBNP (37 kDa), and 2 peaks of BNP immunoreactivity, a major peak (37 kDa) for proBNP, and a minor peak (4 kDa) for BNP. In patient plasma, the molar concentration of NT-proBNP was almost 10 times that of proBNP. The mean proBNP:BNP ratio in patient plasma was 6.3, ranging from 1.8 to 10.8.
Conclusions: ProBNP is the major BNP-immunoreactive form in human blood. The proBNP:BNP ratio in plasma samples is dependent on the methods used for sample handling and for the measurement of the peptides.
The following articles in journals at HighWire Press have cited this article:
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  A. G. Semenov, A. B. Postnikov, N. N. Tamm, K. R. Seferian, N. S. Karpova, M. N. Bloshchitsyna, E. V. Koshkina, M. I. Krasnoselsky, D. V. Serebryanaya, and A. G. Katrukha Processing of Pro-Brain Natriuretic Peptide Is Suppressed by O-Glycosylation in the Region Close to the Cleavage Site Clin. Chem., March 1, 2009; 55(3): 489 - 498. [Abstract] [Full Text] [PDF]
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 P. Lourenco, J. P. Araujo, A. Azevedo, A. Ferreira, and P. Bettencourt The cyclic guanosine monophosphate/B-type natriuretic peptide ratio and mortality in advanced heart failure Eur J Heart Fail, February 1, 2009; 11(2): 185 - 190. [Abstract] [Full Text] [PDF]
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 S. Vickery, M. C. Webb, C. P. Price, R. I. John, N. A. Abbas, and E. J. Lamb Prognostic value of cardiac biomarkers for death in a non-dialysis chronic kidney disease population Nephrol. Dial. Transplant., November 1, 2008; 23(11): 3546 - 3553. [Abstract] [Full Text] [PDF]
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 E. E. Niederkofler, U. A. Kiernan, J. O'Rear, S. Menon, S. Saghir, A. A. Protter, R. W. Nelson, and U. Schellenberger Detection of Endogenous B-Type Natriuretic Peptide at Very Low Concentrations in Patients With Heart Failure Circ Heart Fail, November 1, 2008; 1(4): 258 - 264. [Abstract] [Full Text] [PDF]
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J. P. Goetze, U. Dahlstrom, J. F. Rehfeld, and U. Alehagen Impact of Epitope Specificity and Precursor Maturation in Pro-B-Type Natriuretic Peptide Measurement Clin. Chem., November 1, 2008; 54(11): 1780 - 1787. [Abstract] [Full Text] [PDF]
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N. N. Tamm, K. R. Seferian, A. G. Semenov, K. S. Mukharyamova, E. V. Koshkina, M. I. Krasnoselsky, A. B. Postnikov, D. V. Serebryanaya, F. S. Apple, M. M. Murakami, et al. Novel Immunoassay for Quantification of Brain Natriuretic Peptide and Its Precursor in Human Blood Clin. Chem., September 1, 2008; 54(9): 1511 - 1518. [Abstract] [Full Text] [PDF]
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A. Hammerer-Lercher, B. Halfinger, B. Sarg, J. Mair, B. Puschendorf, A. Griesmacher, N. A. Guzman, and H. H. Lindner Analysis of Circulating Forms of proBNP and NT-proBNP in Patients with Severe Heart Failure Clin. Chem., May 1, 2008; 54(5): 858 - 865. [Abstract] [Full Text] [PDF]
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K. R. Seferian, N. N. Tamm, A. G. Semenov, A. A. Tolstaya, E. V. Koshkina, M. I. Krasnoselsky, A. B. Postnikov, D. V. Serebryanaya, F. S. Apple, M. M. Murakami, et al. Immunodetection of Glycosylated NT-proBNP Circulating in Human Blood Clin. Chem., May 1, 2008; 54(5): 866 - 873. [Abstract] [Full Text] [PDF]
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 G. Szinnai, N. G. Morgenthaler, K. Berneis, J. Struck, B. Muller, U. Keller, and M. Christ-Crain Changes in Plasma Copeptin, the C-Terminal Portion of Arginine Vasopressin during Water Deprivation and Excess in Healthy Subjects J. Clin. Endocrinol. Metab., October 1, 2007; 92(10): 3973 - 3978. [Abstract] [Full Text] [PDF]
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 F. S. Apple, A. H.B. Wu, A. S. Jaffe, M. Panteghini, R. H. Christenson, NACB COMMITTEE MEMBERS, R. H. Christenson, F. S. Apple, C. P. Cannon, G. Francis, et al. National Academy of Clinical Biochemistry and IFCC Committee for Standardization of Markers of Cardiac Damage Laboratory Medicine Practice Guidelines: Analytical Issues for Biomarkers of Heart Failure Circulation, July 31, 2007; 116(5): e95 - e98. [Full Text] [PDF]
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R. L. Gundry and J. E. Van Eyk Unraveling the Complexity of Circulating Forms of Brain Natriuretic Peptide Clin. Chem., July 1, 2007; 53(7): 1181 - 1182. [Full Text] [PDF]
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