Direct chemiluminescent enzyme immunoassay for atrial natriuretic peptide in mammalian plasma using a PEGylated antibody

Nagai, Chiaki; Minamino, Naoto

doi:10.1016/j.ab.2014.05.022

Cited by 15 publications

(18 citation statements)

References 25 publications

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“…Therefore, this DCM model was appropriate to identify new staging biomarkers for DCM. Since plasma levels of well‐known heart failure biomarkers, ANP and BNP, were elevated before 12 wk and maintained at high levels up to 24 wk (ANP, Table and Supporting Information Figure 1A; BNP, Supporting Information Figure 1B), we planned to identify biomarkers sustainably increasing during the course of DCM progression . This is because these biomarkers were expected to be more promising and appropriate as staging biomarkers in the clinical setting.…”

Section: Discussionmentioning

confidence: 99%

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Multiomics approach to identify novel biomarkers for dilated cardiomyopathy: Proteome and transcriptome analyses of 4C30 dilated cardiomyopathy mouse model

et al. 2016

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Dilated cardiomyopathy (DCM) is an intractable disease, without any radical treatment option other than cardiac transplantation. Additionally, biomarkers to determine progressive staging are not yet available. Irrespective of the diversity of causative gene mutations, the phenotype of DCM is rather common. Therefore, it is plausible to determine DCM staging in terms of variations in protein and mRNA levels. In this study, we performed proteome and transcriptome analysis of the left ventricle of 4C30 DCM model mice showing mild and severe phenotypes at 12 and 24 weeks (wk) after birth, respectively. Proteomic analyses results showed 109 proteins that increased and 133 others that decreased among 1874 detected proteins. We selected biomarker candidates by confirming consistent alterations in protein levels at 12 and 24 wk, and mRNA levels at 12 wk, and narrowed these down based on the requirement that they should be detectable in blood. Finally, we selected six biomarker candidates based on sustained or augmented alteration at 24 wk and confirmed their definite alterations in the left ventricle by quantitative polymerase chain reaction, western blot analysis, and multiple reaction monitoring (MRM). To assess the validity of this strategy, we measured plasma concentrations of the six candidates by MRM method and identified two proteins (FTL1 and GRP78) that demonstrated significant elevation in the 4C30 mice plasma. Taken together, a multiomics strategy comparing tissue expression levels of proteins and mRNAs between diseased and control groups, with appropriate confirmation, is a promising approach for the discovery of new biomarkers. © 2016 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 106: 491-502, 2016.

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Section: Discussionmentioning

confidence: 99%

“…Plasma ANP concentration was measured by direct chemiluminescent enzyme immunoassay as previously described, using tenfold diluted plasma.…”

Section: Methodsmentioning

confidence: 99%

Multiomics approach to identify novel biomarkers for dilated cardiomyopathy: Proteome and transcriptome analyses of 4C30 dilated cardiomyopathy mouse model

et al. 2016

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“…After centrifugation at 1,500 g for 20 minutes at 4°C, plasma samples were collected and stored at -80°C until use. Plasma OSTN concentrations were measured by sandwich chemiluminescence enzyme immunoassay (CLEIA), modifying the previously reported protocol for mouse ANP CLEIA (31). Anti-mouse OSTN C-terminus-directed antibody (anti-mMUS1G2) was raised in rabbits against immune complexes conjugated to a custom-synthesized C-terminal octadecapeptide (YGIPMDRIGRNRLSSSRG, N-tyrosinyl mouse preproosteocrin [114 to 130 amino acids]) with the keyhole limpet hemocyanin.…”

Section: Author Contributionsmentioning

confidence: 99%

Circulating osteocrin stimulates bone growth by limiting C-type natriuretic peptide clearance

Kanai¹,

Yasoda²,

Mori³

et al. 2017

Journal of Clinical Investigation

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“…Recombinant human proANP with an N-terminal FLAG tag (FLAG-proANP) was expressed in silkworms and purified using an anti-FLAG affinity column and reversephase (RP)-HPLC as described below. α-ANP fragment peptides were prepared by reduction, carboxymethylation, and subsequent digestion of α-ANP with endoproteinase Asp-N or trypsin (Promega), as described previously (23).…”

Section: Productsmentioning

confidence: 99%

“…Competitive RIAs for epitope analysis of #32-3 and #95-5 were performed as described previously (23). Briefly, 100 μL each of the sample, Ab, and 125 I-labeled tracer solution was mixed and incubated at 4°C for 40 h. The radioactivity of the pellet was quantified using a γ counter (ARC-1000M; Aloka).…”

Section: Preparation and Characterization Of Antibodiesmentioning

confidence: 99%

Novel Chemiluminescent Enzyme Immunoassays for Individual Quantification of 3 Endogenous Molecular Forms of Atrial Natriuretic Peptide in Human Plasma

Nagai‐Okatani

Kangawa

Takashio

et al. 2016

The Journal of Applied Laboratory Medicine

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Background Atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) are cardiac peptide hormones with pivotal roles in maintaining cardiovascular homeostasis. BNP and its precursor fragment are accepted as gold standard markers for heart failure (HF). Human ANP is present in the atria of the heart and plasma as 3 endogenous molecular forms designated α-ANP, β-ANP, and proANP. A previous study indicated that the ratios of these 3 ANP forms are altered in the plasma of HF patients. The purpose of our study was to establish immunoassays for quantifying the individual ANP forms to collect clinical information. Methods We developed 3 plate-based chemiluminescent enzyme immunoassays (CLEIAs) for measuring total ANP (i.e., sum of α-ANP, β-ANP, and proANP), β-ANP, and proANP levels. To minimize background signals, we added single-step PEGylation targeting the immobilized antibody in the conventional plate-based sandwich CLEIA procedure. Results CLEIAs with PEGylation showed sensitivity, specificity, reproducibility, and accuracy satisfying clinical requirements. Two of the CLEIAs enabled direct measurement in plasma samples. During treatments, acute decompensated HF patients exhibited marked decreases in plasma β-ANP levels but moderate decreases in plasma proANP level. The plasma ratios of α-ANP/total ANP and proANP/total ANP in acute decompensated HF patients were maintained, whereas the β-ANP/total ANP ratio was significantly decreased at discharge. Conclusions The combination of the 3 CLEIAs enabled accurate quantification of α-ANP, β-ANP, and proANP, even in plasma samples, and indicated the potential of β-ANP and proANP as circulating biomarkers for HF, with different characteristics from that of BNP.

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Direct chemiluminescent enzyme immunoassay for atrial natriuretic peptide in mammalian plasma using a PEGylated antibody

Cited by 15 publications

References 25 publications

Multiomics approach to identify novel biomarkers for dilated cardiomyopathy: Proteome and transcriptome analyses of 4C30 dilated cardiomyopathy mouse model

Multiomics approach to identify novel biomarkers for dilated cardiomyopathy: Proteome and transcriptome analyses of 4C30 dilated cardiomyopathy mouse model

Circulating osteocrin stimulates bone growth by limiting C-type natriuretic peptide clearance

Novel Chemiluminescent Enzyme Immunoassays for Individual Quantification of 3 Endogenous Molecular Forms of Atrial Natriuretic Peptide in Human Plasma

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