title: Discriminatory proteomic biomarker analysis identifies free hemoglobin in the cerebrospinal fluid of women with severe preeclampsia.
abstract: OBJECTIVE
  Preeclampsia is an idiopathic multisystem disorder specific to human pregnancy. This study used proteomic analysis of cerebrospinal fluid (CSF) to identify protein biomarkers characteristic of preeclampsia and related to its severity.
  
  
  METHODS
  CSF was collected from women diagnosed clinically with severe preeclampsia (sPE: n = 7), mild preeclampsia (mPE: n = 8), and normotensive controls (CRL: n = 8). Samples were subjected to proteomic analysis using surface-enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectroscopy. A discriminative proteomic biomarker profile was extracted by applying Mass Restricted analysis, and a Preeclampsia Proteomic Biomarker (PPB) score developed based on the presence or absence of four discriminatory protein peaks in individual CSF SELDI tracings. In-gel tryptic digests, Western blot analysis, on-chip immunoassays, ELISA, and spectral analysis were used to identify the biomarkers composing the PPB score.
  
  
  RESULTS
  PPB score distinguished patients with a clinical diagnosis of sPE from mPE and CRLs. (PPB median [range]: sPE: 4 [0-4] vs mPE: 1 [0-1] vs CRL: 0 [0-0]; P < 0.001). PPB scores were unaffected by parity, magnesium seizure prophylaxis, CSF leukocyte counts, and total protein content. Proteomic identification techniques matched the discriminatory protein peaks to the alpha- and beta-hemoglobin chains. ELISA confirmed that women diagnosed clinically with sPE had significantly higher CSF hemoglobin concentrations than women with mPE or CRL (median [range]: sPE: 6.6 [0.0-10.3] microg/mL vs mPE: 0 [0-1.3] microg/mL vs CRL: 0 [0-0] microg/mL; P < 0.001).
  
  
  CONCLUSIONS
  Proteomic analysis of CSF can accurately distinguish sPE from both mPE and CRL. Patients with sPE have nanomolar amounts of free hemoglobin in their CSF. Further studies are needed to confirm these observations and determine their physiologic implications.
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, New Haven, CT, USA.
authors.forename: Errol R
authors.initials: ER
authors.lastname: Norwitz
authors.forename: Lawrence C
authors.initials: LC
authors.lastname: Tsen
authors.forename: Joong Shin
authors.initials: JS
authors.lastname: Park
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Fitzpatrick
authors.forename: David M
authors.initials: DM
authors.lastname: Dorfman
authors.forename: George R
authors.initials: GR
authors.lastname: Saade
authors.forename: Catalin S
authors.initials: CS
authors.lastname: Buhimschi
authors.forename: Irina A
authors.initials: IA
authors.lastname: Buhimschi
mesh_terms: D015415:Biomarkers / Q000032:analysis; D005260:Female; D006454:Hemoglobins / Q000134:cerebrospinal fluid*; D006801:Humans; D013058:Mass Spectrometry; D011225:Pre-Eclampsia / Q000134:cerebrospinal fluid*; D011247:Pregnancy; D040081:Protein Array Analysis; D040901:Proteomics
chemical_list: D015415:Biomarkers; D006454:Hemoglobins