Competitive ELISA for N-terminal pro-brain natriuretic peptide (NT-proBNP) determination in human plasma

Colom, Gloria; Salvador, J.‐Pablo; Acosta, Gerardo; Alberício, Fernando; Royo, Míriam; Marco, M.‐Pilar

doi:10.1039/d0an00650e

Cited by 12 publications

(8 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Key elements for setting up a multiplexed diagnostic kit are the availability of immunoreagents, appropriately labeled to provide a signal in the presence of the biomarker target, and the solid support in which the immunochemical reaction is going to take place. Recently, we have reported the development of antibodies and an ELISA for NT-proBNP biomarker using competitive immunoassay format [ 25 ]. Moreover, previously, we developed antibodies against cTnI and use them on a sandwich configuration to develop an electrochemical immunosensor [ 24 ].…”

Section: Resultsmentioning

confidence: 99%

“…In order to standardize detection of the immunochemical reaction, the antibodies were biotinylated. Thus, the proper bioconjugates should be prepared as a competitor for NT-proBNP [ 25 ] and biotinylation of detection antibodies. The biotinylation was required in order to homogenize the detection antibodies (CRP, CysC, and h-FABP) that were from different immunized animals and avoid cross-reactivity for the capture antibodies.…”

Section: Resultsmentioning

confidence: 99%

“…All salts were provided by Merck (Darmstadt, Germany). The procedure for the preparation of NB1-SIA-BSA conjugate and for the production of As 251 is already described [ 25 ]. Goat Anti-Rabbit IgG H&L TRITC conjugate from Abcam Plc (Cambridge, UK).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

A multiplexed immunochemical microarray for the determination of cardiovascular disease biomarkers

Colom,

Hernandez-Albors,

Barallat

et al. 2023

Microchim Acta

Self Cite

View full text Add to dashboard Cite

A fluorescence antibody microarray has been developed for the determination of relevant cardiovascular disease biomarkers for the analysis of human plasma samples. Recording characteristic protein molecular fingerprints to assess individual’s states of health could allow diagnosis to go beyond the simple identification of the disease, providing information on its stage or prognosis. Precisely, cardiovascular diseases (CVDs) are complex disorders which involve different degenerative processes encompassing a collection of biomarkers related to disease progression or stage. The novel approach that we propose is a fluorescent microarray chip has been developed accomplishing simultaneous determination of the most significant cardiac biomarkers in plasma aiming to determine the CVD status stage of the patient. As proof of concept, we have chosen five relevant biomarkers, C-reactive protein (CRP) as biomarker of inflammation, cystatin C (CysC) as biomarker of renal failure that is directly related with heart failure, cardiac troponin I (cTnI) as already established biomarker for cardiac damage, heart fatty acid binding protein as biomarker of ischemia (H-FABP), and finally, NT-proBNP (N-terminal pro-brain natriuretic peptide), a well-established heart failure biomarker. After the optimization of the multiplexed microarray, the assay allowed the simultaneous determination of 5 biomarkers in a buffer solution reaching LODs of 15 ± 5, 3 ± 1, 24 ± 3, 25 ± 3, and 3 ± 1 ng mL−1, for CRP, CysC, H-FABP, cTnI, and NT-proBNP, respectively. After solving the matrix effect, and demonstrating the accuracy for each biomarker, the chip was able to determine 24 samples per microarray chip. Then, the microarray has been used on a small pilot clinical study with 29 plasma samples from clinical patients which suffered different CVD and other related disorders. Results show the superior capability of the chip to provide clinical information related to the disease in terms of turnaround time (1 h 30 min total assay and measurement) and amount of information delivered in respect to reference technologies used in hospital laboratories (clinical analyzers). Despite the failure to detect c-TnI at the reported threshold, the microarray technology could be a powerful approach to diagnose the cardiovascular disease at early stage, monitor its progress, and eventually providing information about an eminent potential risk of suffering a myocardial infarction. The microarray chip here reported could be the starting point for achieving powerful multiplexed diagnostic technologies for the diagnosis of CVDs or any other pathology for which biomarkers have been identified at different stages of the disease. Graphical Abstract

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

A multiplexed immunochemical microarray for the determination of cardiovascular disease biomarkers

Colom,

Hernandez-Albors,

Barallat

et al. 2023

Microchim Acta

Self Cite

View full text Add to dashboard Cite

show abstract

“…The usage of earlier techniques, such as enzyme-linked immunosorbent assay (ELISA) and immunochromatographic test, is constrained by the need for basic equipment, like microplate readers and strip readers. [6][7][8][9][10][11][12] The latter assays include electrochemical biosensors, photoelectrochemical biosensors, surface plasma resonance biosensors, electrochemiluminescence biosensors, and micro-uidic immunoassay biosensors, among others. [13][14][15][16][17][18][19][20][21][22] Large instruments are necessary for these delicate biosensors, which are expensive, uncomfortable for users, and need trained technicians.…”

Section: Introductionmentioning

confidence: 99%

Shape dependency of gold nanorods through TMB²⁺-mediated etching for the visual detection of NT-proBNP

et al. 2023

View full text Add to dashboard Cite

show abstract

“…Studies show that in addition to vitronectin, hs-CRP, like other risk factors, plays a role in this process (11-13). Also, proBNP is a cardiac natriuretic peptide synthesized and secreted in response to increased ventricular wall stress in the ventricular myocardium (20).…”

Section: Introductionmentioning

confidence: 99%

The relationship of hs-CRP, vitronectin and NT-proBNP serum levels with the extent and severity of cardiac complications in patients with organophosphate pesticide poisoning

You-gui

Sun

Han

et al. 2022

Cell Mol Biol (Noisy-le-grand)

View full text Add to dashboard Cite

Acute organophosphate poisoning kills tens of thousands of people annually around the world. These substances are widely used as insecticides in homes, industry, and agricultural environments. Due to the ease of access, they can cause accidental or intentional risks of exposure through the skin or respiratory contact. This study aimed to evaluate the serum levels of hs-CRP, Vitronectin, and NT-proBNP and their relationship with the extent and severity of cardiac complications in patients with organophosphate pesticide poisoning. In this descriptive-comparative study, 160 patients were studied with acute organophosphate poisoning. Also, for better comparison, 40 healthy individuals participated in this study. Diagnosis of organophosphate poisoning was based on clinical findings of serum butyrylcholinesterase levels. The hs-CRP measurement was performed by an autoanalyzer (Abbott, model Alcyon 300, USA) with the ELISA hs-CRP kit (The apDia Company, Belgium). Vitronectin (VN) measurements were performed by ELISA method and Glory science human VN kit with Catalog No: 11668. NT-ProBNP serum levels were analyzed by ProBNP assay kit (Roche, Germany) by ECLIA method using Elecsys 2010 Analyzer. The most important variables studied in this study were the electrical activity and conduction system of the heart, PR distance, QTC interval, and T-wave changes. In this study, most of the patients were women and girls (60.78%). The highest percentage of organophosphate poisoning was in the age group of 15-24 years (37.25%). In most cases (78.43%), poisoning was intentional or suicidal. Evaluation of electrophysiological abnormalities of the heart showed that 89 patients (55.62%) had long QTC interval (>450 msec), 43 cases (26.87%) had possible long QTC (431-450 msec), and 28 cases (17.5%) had normal QTC (<430 msec). Only 9.37% of cases (n = 15) showed an increase in P-R distance, which is characteristic of the first-degree ventricular atrial block. Sinus bradycardia occurred in 57 cases (35.62%) and sinus tachycardia in 43 cases (26.87%); in 60 cases (37.5%), the pulse rate was normal. Smooth T-wave changes were observed in 9.8% of patients and reverse T-wave was observed in 17.6%. A long T-wave was not reported in any case. In only two cases (1.25%) was grade 1 ventricular atrial block and grade 2 and 3 blocks were not observed. In general, there was a significant difference in the hs-CRP, vitronectin, and NT-proBNP serum levels between the patient and control groups in all studied variables. These parameters were also related to the extent and severity of the disease.

show abstract

Competitive ELISA for N-terminal pro-brain natriuretic peptide (NT-proBNP) determination in human plasma

Abstract: Brain natriuretic peptides (BNPs) are well established cardiovascular disease (CVD) biomarkers, which are released from the heart after ventricular wall stress. Particularly, the N-terminal proBNP (NT-proBNP) is a 76 aa...

Cited by 12 publications

References 24 publications

A multiplexed immunochemical microarray for the determination of cardiovascular disease biomarkers

A multiplexed immunochemical microarray for the determination of cardiovascular disease biomarkers

Shape dependency of gold nanorods through TMB²⁺-mediated etching for the visual detection of NT-proBNP

The relationship of hs-CRP, vitronectin and NT-proBNP serum levels with the extent and severity of cardiac complications in patients with organophosphate pesticide poisoning

Contact Info

Product

Resources

About

Competitive ELISA for N-terminal pro-brain natriuretic peptide (NT-proBNP) determination in human plasma

Abstract: Brain natriuretic peptides (BNPs) are well established cardiovascular disease (CVD) biomarkers, which are released from the heart after ventricular wall stress. Particularly, the N-terminal proBNP (NT-proBNP) is a 76 aa...

Cited by 12 publications

References 24 publications

A multiplexed immunochemical microarray for the determination of cardiovascular disease biomarkers

A multiplexed immunochemical microarray for the determination of cardiovascular disease biomarkers

Shape dependency of gold nanorods through TMB2+-mediated etching for the visual detection of NT-proBNP

The relationship of hs-CRP, vitronectin and NT-proBNP serum levels with the extent and severity of cardiac complications in patients with organophosphate pesticide poisoning

Contact Info

Product

Resources

About

Shape dependency of gold nanorods through TMB²⁺-mediated etching for the visual detection of NT-proBNP