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 | |
| State | Published - Jan 2011 |
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Keywords
- bufodienolides
- cardiac glycosides
- diagnostic
- hypertension
- immunoassay
- marinobufagenin
ASJC Scopus subject areas
- Immunology
- Immunology and Allergy
- Clinical Biochemistry
- Medical Laboratory Technology
Cite this
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
}
TY - JOUR
T1 - A chemifluorescent immunoassay for the determination of marinobufagenin in body fluids
AU - Abi-Ghanem, Daad
AU - Lai, Xinzhong
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.
PY - 2011/1
Y1 - 2011/1
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.
KW - bufodienolides
KW - cardiac glycosides
KW - diagnostic
KW - hypertension
KW - immunoassay
KW - marinobufagenin
UR - http://www.scopus.com/inward/record.url?scp=79251471785&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79251471785&partnerID=8YFLogxK
U2 - 10.1080/15321819.2010.538107
DO - 10.1080/15321819.2010.538107
M3 - Article
C2 - 21253968
AN - SCOPUS:79251471785
VL - 32
SP - 31
EP - 46
JO - Journal of Immunoassay and Immunochemistry
JF - Journal of Immunoassay and Immunochemistry
SN - 1532-1819
IS - 1
ER -