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Ann Clin Biochem. 2016 Jan;53(Pt 1):85-96. doi: 10.1177/0004563215583697. Epub 2015 Apr 2.

Simple quantitation for potential serum disease biomarker peptides, primarily identified by a peptidomics approach in the serum with hypertensive disorders of pregnancy.

Author information

1
Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Chiba, Japan Department of Obstetrics and Gynecology, Juntendo University Graduate School of Medicine, Tokyo, Japan.
2
Membrane Protein and Ligand Analysis Center, Protosera Inc., Hyogo, Japan.
3
Department of Health Information Management, Yamagata Saisei Hospital, Yamagata, Japan.
4
Department of Obstetrics and Gynecology, Yamagata Saisei Hospital, Yamagata, Japan.
5
Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Chiba, Japan.
6
Department of Obstetrics and Gynecology, Juntendo University Graduate School of Medicine, Tokyo, Japan.
7
Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Chiba, Japan Department of Obstetrics and Gynecology, Juntendo University Graduate School of Medicine, Tokyo, Japan yaraki@juntendo.ac.jp.

Abstract

BACKGROUND:

We previously reported peptide candidates of disease biomarkers for pregnancy-induced hypertension syndrome using a novel peptidomic analytical method, BLOTCHIP®-MS. The aim of this study was to establish a sandwich enzyme-linked immunosorbent assay system for quantitation of such peptides and to validate their usefulness as disease biomarkers of pregnancy-induced hypertension syndrome including gestational hypertension/pre-eclampsia.

METHODS:

We focused on three peptide fragments, kininogen-1439-456 (PDA039), kininogen-1438-456 (PDA044) and cysteinyl α2-HS-glycoprotein341-367 (PDA071). Using polyclonal antibodies specific for each peptide, suitable conditions for the sandwich enzyme-linked immunosorbent assay system were investigated. The quantitative enzyme-linked immunosorbent assay values were confirmed by quantitative matrix assisted laser desorption/ionization time-of-flight MS analyses. Using the established enzyme-linked immunosorbent assay systems, serum samples from gestational hypertension/pre-eclampsia patients and paired serum samples from healthy pregnant females were analysed.

RESULTS:

The optimum sandwich enzyme-linked immunosorbent assay conditions for PDA039/044 quantitation were developed. Quantitation of PDA071 by enzyme-linked immunosorbent assay failed, presumably due to issues with polyclonal antibody specificity for the native peptide. Bland-Altman plots showed a satisfactory correlation between the serum PDA039/044 concentration by enzyme-linked immunosorbent assay and that by quantitative MS analysis. Although the PDA044 concentration showed no significant change during pregnancy, including gestational hypertension/pre-eclampsia patients, the serum PDA039 concentration was significantly increased (P < 0.0001) in the patients.

CONCLUSIONS:

The simple quantitation technology for PDA039 by enzyme-linked immunosorbent assay was established for the first time. PDA039 confirmed its clinical utility as a disease biomarker for gestational hypertension/pre-eclampsia by the enzyme-linked immunosorbent assay system using clinical samples. The information provided from the present study would be a new valuable addition in the field of gestational hypertension/pre-eclampsia research.

KEYWORDS:

Sandwich enzyme-linked immunosorbent assays; hypertensive disorders of pregnancy; peptidomics; serum biomarkers

PMID:
25838414
DOI:
10.1177/0004563215583697
[Indexed for MEDLINE]

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