A chemifluorescent immunoassay for the determination of marinobufagenin in body fluids

Daad Abi-Ghanem, Xinzhong Lai, Luc R. Berghman, Darijana Horvat, Jing Li, Daniel Romo, Mohammad N. Uddin, Yoshiaki Kamano, Toshihiko Nogawa, Jun-Ping Xu, George Pettit, Jules B. Puschett

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

We describe here the development of a chemifluorescent competitive enzyme-linked immunosorbent assay (ELISA) that quantifies marinobufagenin (MBG) levels in biological fluids. Based on a polyclonal antibody raised against a novel MBG-bovine serum albumin conjugate, this assay achieved an MBG detection limit of less than 9pg/mL. MBG levels in various rat urine and serum samples were effectively determined using this methodology. Interassay variability averaged 9.8%, while intra-assay variability averaged 1.9 and 2.5% in representative serum and urine samples, respectively. Recovery of exogenously added MBG averaged 106%, and parallelism data further established the accuracy of the assay. Employment of this assay to detect MBG abnormalities represents a powerful tool for the possible diagnosis, prevention and management of human hypertensive states, particularly preeclampsia.

Original languageEnglish (US)
Pages (from-to)31-46
Number of pages16
JournalJournal of Immunoassay and Immunochemistry
Volume32
Issue number1
DOIs
StatePublished - Jan 2011

Keywords

  • bufodienolides
  • cardiac glycosides
  • diagnostic
  • hypertension
  • immunoassay
  • marinobufagenin

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Clinical Biochemistry
  • Medical Laboratory Technology

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