This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Buhimschi, I. A. Articles by Ratner, E. Search for Related Content PubMed PubMed Citation Articles by Buhimschi, I. A. Articles by Ratner, E.

Proteomic but Not Enzyme-Linked Immunosorbent Assay Technology Detects Amniotic Fluid Monomeric Calgranulins from Their Complexed Calprotectin Form

Department of Obstetrics, Gynecology, and Reproductive Science, Yale University, 333 Cedar Street, New Haven, Connecticut 06520-8063,¹ Department of Obstetrics, Gynecology, and Reproductive Science, University of Maryland at Baltimore, Baltimore, Maryland 21044,² Advanced Life Science Institute, Wako, Saitama, Japan³

Received 21 February 2005/ Returned for modification 20 April 2005/ Accepted 29 April 2005

Four proteomic biomarkers (human neutrophil peptide 1 [HNP1], HNP2 [defensins], calgranulin C [Cal-C], and Cal-A) characterize the fingerprint of intra-amniotic inflammation (IAI). We compared proteomic technology using surfaced-enhanced laser desorption-ionization-time of flight (SELDI-TOF) mass spectrometry to enzyme-linked immunosorbent assay (ELISA) for detection of these biomarkers. Amniocentesis was performed on 48 women enrolled in two groups: those with intact membranes (n = 27; gestational age [GA], 26.0 ± 0.8 weeks) and those with preterm premature rupture of the membranes (PPROM; n = 21; GA, 28.4 ± 0.9 weeks). Paired abdominal amniotic fluids (aAFs)-vaginal AFs (vAFs) were analyzed in PPROM women. Quantitative aspects of HNP1-3, Cal-C, Cal-A, and calprotectin (a complex of Cal-A with Cal-B) were assessed by ELISA. SELDI-TOF mass spectrometry tracings from 16/48 (33.3%) aAFs and 13/17 (88.2%) vAFs were consistent with IAI (three or four biomarkers present). IAI (by SELDI-TOF mass spectrometry) was associated with increased HNP1-3 and Cal-C measured by ELISA. However, immunoassays detected Cal-A in only 4 of the AFs even though its specific SELDI-TOF mass spectrometry peak was identified in 19/48 AFs. Calprotectin immunoreactivity was decreased in AFs retrieved from women with IAI (P = 0.01). In conclusion, IAI is associated with increased HNP1-3 levels. In the absence of isoform-specific ELISAs, mass spectrometry remains the only way to discriminate the HNP biomarker isoforms. Monomeric Cal-A is not reliably estimated by specific ELISA as it binds to Cal-B to form the calprotectin complex. Cal-C was reliably measured by SELDI-TOF mass spectrometry or specific ELISA.

This article has been cited by other articles:

• Klein, L. L., Jonscher, K. R., Heerwagen, M. J., Gibbs, R. S., McManaman, J. L. (2008). Shotgun Proteomic Analysis of Vaginal Fluid From Women in Late Pregnancy. Reproductive Sciences 15: 263-273 [Abstract]