Circulating Fragments of N-Terminal Pro-B-Type Natriuretic Peptides in Plasma of Heart Failure Patients

Foo, Jared Yong Yang, Wan, Yunxia, Schulz, Benjamin L., Kostner, Karam, Atherton, John, Cooper-White, Justin, Dimeski, Goce and Punyadeera, Chamindie (2013) Circulating Fragments of N-Terminal Pro-B-Type Natriuretic Peptides in Plasma of Heart Failure Patients. Clinical Chemistry, 59 10: 1523-1531. doi:10.1373/clinchem.2012.200204


Author Foo, Jared Yong Yang
Wan, Yunxia
Schulz, Benjamin L.
Kostner, Karam
Atherton, John
Cooper-White, Justin
Dimeski, Goce
Punyadeera, Chamindie
Title Circulating Fragments of N-Terminal Pro-B-Type Natriuretic Peptides in Plasma of Heart Failure Patients
Journal name Clinical Chemistry   Check publisher's open access policy
ISSN 0009-9147
1530-8561
Publication date 2013-10-01
Year available 2013
Sub-type Article (original research)
DOI 10.1373/clinchem.2012.200204
Open Access Status Not Open Access
Volume 59
Issue 10
Start page 1523
End page 1531
Total pages 9
Place of publication Washington, United States
Publisher American Association for Clinical Chemistry
Language eng
Formatted abstract
BACKGROUND: The use of nonstandardized N-terminal pro–B-type natriuretic peptide (NT-proBNP) assays can contribute to the misdiagnosis of heart failure (HF). Moreover, there is yet to be established a common consensus regarding the circulating forms of NT-proBNP being used in current assays. We aimed to characterize and quantify the various forms of NT-proBNP in the circulation of HF patients.

METHODS: Plasma samples were collected from HF patients (n = 20) at rest and stored at −80 °C. NT-proBNP was enriched from HF patient plasma by use of immunoprecipitation followed by mass spectrometric analysis. Customized homogeneous sandwich AlphaLISA® immunoassays were developed and validated to quantify 6 fragments of NT-proBNP.

RESULTS: Mass spectrometry identified the presence of several N- and C-terminally processed forms of circulating NT-proBNP, with physiological proteolysis between Pro2-Leu3, Leu3-Gly4, Pro6-Gly7, and Pro75-Arg76. Consistent with this result, AlphaLISA immunoassays demonstrated that antibodies targeting the extreme N or C termini measured a low apparent concentration of circulating NT-proBNP. The apparent circulating NT-proBNP concentration was increased with antibodies targeting nonglycosylated and nonterminal epitopes (P < 0.05).

CONCLUSIONS: In plasma collected from HF patients, immunoreactive NT-proBNP was present as multiple N- and C-terminally truncated fragments of the full length NT-proBNP molecule. Immunodetection of NT-proBNP was significantly improved with the use of antibodies that did not target these terminal regions. These findings support the development of a next generation NT-proBNP assay targeting nonterminal epitopes as well as avoiding the central glycosylated region of this molecule.
Keyword Saliva
Assay
Corin
State
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
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