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

doi:10.1080/15321819.2010.538107

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 journal › Article › peer-review

7 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 language	English (US)
Pages (from-to)	31-46
Number of pages	16
Journal	Journal of Immunoassay and Immunochemistry
Volume	32
Issue number	1
DOIs	https://doi.org/10.1080/15321819.2010.538107
State	Published - 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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10.1080/15321819.2010.538107

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A chemifluorescent immunoassay for the determination of marinobufagenin in body fluids. / Abi-Ghanem, Daad; Lai, Xinzhong; Berghman, Luc R.; Horvat, Darijana; Li, Jing; Romo, Daniel; Uddin, Mohammad N.; Kamano, Yoshiaki; Nogawa, Toshihiko; Xu, Jun-Ping; Pettit, George; Puschett, Jules B.

In: Journal of Immunoassay and Immunochemistry, Vol. 32, No. 1, 01.2011, p. 31-46.

Research output: Contribution to journal › Article › peer-review

Abi-Ghanem, Daad ; Lai, Xinzhong ; Berghman, Luc R. ; Horvat, Darijana ; Li, Jing ; Romo, Daniel ; Uddin, Mohammad N. ; Kamano, Yoshiaki ; Nogawa, Toshihiko ; Xu, Jun-Ping ; Pettit, George ; Puschett, Jules B. / A chemifluorescent immunoassay for the determination of marinobufagenin in body fluids. In: Journal of Immunoassay and Immunochemistry. 2011 ; Vol. 32, No. 1. pp. 31-46.

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title = "A chemifluorescent immunoassay for the determination of marinobufagenin in body fluids",

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.",

keywords = "bufodienolides, cardiac glycosides, diagnostic, hypertension, immunoassay, marinobufagenin",

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

note = "Funding Information: This work was supported (in part) by grants-in-aid from the Joe W. and Dorothy Dorsett Brown Foundation, New Orleans, LA, Dialysis Clinic, Inc., Nashville, TN, from Scott & White Healthcare, Inc., Temple, TX, the Texas A&M Health Science Center College of Medicine, College Station, TX (to JBP), and by grants from the USDA (grant 2008-35204-04554 to LRB), from the National Institutes of Health (RO1 GM086307 to DR), grants R01 CA 90441-01-04, 5R01, and CA090441-07-08 from the Division of Cancer Treatment and Diagnosis, NCI, DHHS (GRP), and the Welch Foundation (A-1280 to DR). In addition, funding from the College of Science, Texas A&M University (for JL) is gratefully acknowledged. The authors thank Lonnie Doyle for the production of this article.",

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AU - Berghman, Luc R.

AU - Horvat, Darijana

AU - Li, Jing

AU - Romo, Daniel

AU - Uddin, Mohammad N.

AU - Kamano, Yoshiaki

AU - Nogawa, Toshihiko

AU - Xu, Jun-Ping

AU - Pettit, George

AU - Puschett, Jules B.

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N2 - 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.

AB - 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.

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A chemifluorescent immunoassay for the determination of marinobufagenin in body fluids

Abstract

Keywords

ASJC Scopus subject areas

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