Development of a homogeneous immunoassay for the detection of angiotensin I in plasma using AlphaLISA acceptor beads technology

Cauchon, Elizabeth; Liu, Susana; Percival, M. David; Rowland, Steve E.; Xu, Daigen; Binkert, Christoph A.; Strickner, Panja; Falgueyret, Jean‐Pierre

doi:10.1016/j.ab.2009.02.031

Cited by 33 publications

(27 citation statements)

References 13 publications

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“…The AlphaLISA (PerkinElmer) is a homogeneous proximity-based assay in which two types of 200 nm diameter neutral density beads, a donor and an acceptor, are drawn together by the biological interaction of their conjugates [25,26]. Using AlphaLISA, it is possible to measure the relative signal intensity for the total amount of a given glycoprotein using antibody-conjugated beads and that of the glycosylated fraction using lectin-conjugated beads [27].…”

Section: Introductionmentioning

confidence: 99%

Development of an AlphaLISA assay to quantify serum core-fucosylated E-cadherin as a metastatic lung adenocarcinoma biomarker

Wen

Chen

et al. 2012

Journal of Proteomics

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Section: Introductionmentioning

confidence: 99%

Development of an AlphaLISA assay to quantify serum core-fucosylated E-cadherin as a metastatic lung adenocarcinoma biomarker

Wen

Chen

et al. 2012

Journal of Proteomics

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“…In vitro activation of ReninSense by plasma from mice on a low-salt diet (LSD). A: mouse plasma under a normal diet or LSD was harvested and assessed for plasma renin activity (PRA) using a competition-based enzyme immunoassay as previously described (3). Values are means Ϯ SE.…”

Section: In Vitro Assessment Of Reninsense Activation By Renin and Otmentioning

confidence: 99%

A fluorogenic near-infrared imaging agent for quantifying plasma and local tissue renin activity in vivo and ex vivo

Zhang

Preda

Vasquez

et al. 2012

American Journal of Physiology-Renal Physiology

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. A fluorogenic near-infrared imaging agent for quantifying plasma and local tissue renin activity in vivo and ex vivo. Am J Physiol Renal Physiol 303: F593-F603, 2012. First published June 6, 2012; doi:10.1152/ajprenal.00361.2011.-The renin-angiotensin system (RAS) is well studied for its regulation of blood pressure and fluid homeostasis, as well as for increased activity associated with a variety of diseases and conditions, including cardiovascular disease, diabetes, and kidney disease. The enzyme renin cleaves angiotensinogen to form angiotensin I (ANG I), which is further cleaved by angiotensin-converting enzyme to produce ANG II. Although ANG II is the main effector molecule of the RAS, renin is the rate-limiting enzyme, thus playing a pivotal role in regulating RAS activity in hypertension and organ injury processes. Our objective was to develop a near-infrared fluorescent (NIRF) renin-imaging agent for noninvasive in vivo detection of renin activity as a measure of tissue RAS and in vitro plasma renin activity. We synthesized a reninactivatable agent, ReninSense 680 FAST (ReninSense), using a NIRF-quenched substrate derived from angiotensinogen that is cleaved specifically by purified mouse and rat renin enzymes to generate a fluorescent signal. This agent was assessed in vitro, in vivo, and ex vivo to detect and quantify increases in plasma and kidney renin activity in sodium-sensitive inbred C57BL/6 mice maintained on a low dietary sodium and diuretic regimen. Noninvasive in vivo fluorescence molecular tomographic imaging of the ReninSense signal in the kidney detected increased renin activity in the kidneys of hyperreninemic C57BL/6 mice. The agent also effectively detected renin activity in ex vivo kidneys, kidney tissue sections, and plasma samples. This approach could provide a new tool for assessing disorders linked to altered tissue and plasma renin activity and to monitor the efficacy of therapeutic treatments. optical tomography; low-salt diet; hypertension THE RENIN-ANGIOTENSIN SYSTEM (RAS) plays an important role in the regulation of blood volume, electrolyte homeostasis, and systemic vascular resistance, which together affect cardiac output and arterial pressure (11). Renin, a highly specific aspartyl protease, is primarily stored and released from juxtaglomerular (JG) cells within the afferent arterioles of the kidney glomeruli and plays an important regulatory role in RAS function. JG cells can sense a reduction in afferent arteriole pressure in the kidney leading to direct release of renin, and specialized adjacent cells (the macula densa) can also trigger JG renin release upon sensing of decreased plasma sodium chloride in renal tubules or changes in kidney sympathetic nerve signaling. Release of renin into the blood then mediates the first and rate-limiting step of the RAS cascade by cleaving angiotensinogen to generate angiotensin I (ANG I). ANG I is further cleaved by angiotensin-converting enzyme (ACE) to generate bioactive angiotensin II (ANG II), which binds to angiotensin recepto...

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“…This technology provides a facile assay platform with which one can quantify large samples using no-wash microtitration strips in a short amount of time at relatively low cost compared with FMAT and ECL technologies. [19][20][21][22][23][24] Studies have shown that the concentration of free b-hCG differs in different races as well as with respect to weekly median values. [9] In the present study, we used a novel 96-well plate-based AlphaLISA to detect the concentrations of free b-hCG in 4168 serum samples collected from southern Chinese women during 8-20 weeks of gestation.…”

Section: Introductionmentioning

confidence: 99%

Alphalisa for the Determination of Median Levels of the Free Β Subunit of Human Chorionic Gonadotropin in the Serum of Pregnant Women

Zou

Liu

Lin

et al. 2013

Journal of Immunoassay and Immunochemistry

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Measurement of the free β subunit of human chorionic gonadotropin (free β-hCG) in serum is useful for prenatal screening. Concentrations of free β-hCG vary in different races. Conventional assays used for such measurements have limitations. We applied the AlphaLISA to measure levels of free β-hCG in maternal serum during 8-20 weeks of gestation in women from southern China. Two anti-free β-hCG antibodies were used: one was coated on AlphaLISA acceptor beads and the one was biotinylated. The assay also contained donor beads coated with streptavidin. The AlphaLISA assay detection limit was 0.11 ng/mL, and the analytical range was 0.11-200 ng/mL. The intra- and interassay coefficients of variation were 1.32%-2.50% and 3.44%-5.45%, respectively. The correlation with commercial Eu(3+)-labeled free β-HCG-TRFIA assay was good (y = 1.045x + 1.580, r(2) = 0.978). Median levels of free β-hCG in maternal serum at 8-20 weeks gestation were higher in women from southern China compared with those reported in women from other countries. The AlphaLISA for free β-HCG could become the assay of choice for applications in clinical diagnostics. The established median value of free β-HCG is helpful in clinical diagnosis specific for southern Chinese women.

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Development of a homogeneous immunoassay for the detection of angiotensin I in plasma using AlphaLISA acceptor beads technology

Cited by 33 publications

References 13 publications

Development of an AlphaLISA assay to quantify serum core-fucosylated E-cadherin as a metastatic lung adenocarcinoma biomarker

Development of an AlphaLISA assay to quantify serum core-fucosylated E-cadherin as a metastatic lung adenocarcinoma biomarker

A fluorogenic near-infrared imaging agent for quantifying plasma and local tissue renin activity in vivo and ex vivo

Alphalisa for the Determination of Median Levels of the Free Β Subunit of Human Chorionic Gonadotropin in the Serum of Pregnant Women

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