N-terminal Brain Natriuretic Peptide Precursor (NT-proBNP) Employing Magnetic Particle Chemiluminescence with Acridinium Ester and Fluorescence Immuno
Time：2024-02-23 Hits：222
1 Project Background
Excerpted from "Guiding Principles for Technical Review of Registration of Brain Natriuretic Peptide/N-Terminal Brain Natriuretic Peptide Precursor Detection Reagents."
Brain natriuretic peptide (BNP) is a neurohormonal substance with functions akin to atrial natriuretic peptide (ANP). It promotes urination, natriuresis, and resists sympathetic nerve activity. BNP is closely tied to the tissue distribution and amino acid composition of atrial natriuretic peptide, but substantial differences exist in other aspects, categorizing it as a distinct substance named BNP.
Research indicates that BNP in the human body arises from cleaving the C-terminus of the BNP amino acid precursor protein (proBNP). The N-terminus cleavage forms NT-proBNP, an amino-terminal brain natriuretic peptide precursor with 76 amino acids.
The genes of BNP and NT-proBNP differ significantly, primarily manifested in distinct elimination pathways and half-lives within the body. Both are mainly eliminated through the liver and kidneys. BNP undergoes elimination via binding to natriuretic peptide receptor C, neutral endopeptidase, and passive excretion through the kidneys. In contrast, NT-proBNP lacks active clearance mechanisms, relying on passive clearance through glomerular filtration, resulting in a longer detection window.
BNP and NT-proBNP exhibit disparate biological activities. BNP, with a characteristic amino acid ring, possesses important biological activity but has a short half-life of 18-22 minutes. NT-proBNP, with a linear structure and no biological activity, has a longer half-life of approximately 120 minutes, remaining stable and unaffected by factors like body position and daily activities.
Synthesized and secreted in equal molar numbers by left ventricular cardiomyocytes, both BNP and NT-proBNP serve as effective heart failure markers. They independently predict increased ventricular pressure, providing valuable insights in the auxiliary diagnosis, treatment evaluation, and monitoring of congestive heart failure, dyspnea complications, essential hypertension, and cardiovascular-related diseases.
2.Performance
2.1 Chemiluminescence (acridinium ester) Platform
2.1.1 Combination 1
N-terminal pro-brain natriuretic peptide (NT-proBNP) coated carboxyl magnetic beads (Cat. No.: EKY0090L-1mg) + NT-proBNP labeled acridinium ester (Cat. No.: EKY0090M-1mg).
Linear range: 10pg/ml-35000pg/ml.
Stability under thermal acceleration at 37°C for 7 days: luminescence value changes around ±13%.
Enterprise working calibration product (liquid) 7-day stability at 37°C thermal acceleration: luminescence value changes around ±15%.
2.1.2 Combination 2:
NT-proBNP coated carboxyl magnetic beads (Cat. No.: EKY0090P-1mg) + NT-proBNP labeled acridinium ester (Cat. No.: EKY0090Q-1mg).
Linear range: 10pg/ml-35000pg/ml.
2.2 Combination:
NT-proBNP labeled fluorescent microspheres (Cat. No.: DKY0090N-1mg) + NT-proBNP coated scratch film (Cat. No.: DKY0090O-1mg).
Linear range: 50pg/ml-35000pg/ml.
3 Clinical Relevance
3.1 Chemiluminescence (acridinium ester) Platform:
Clinical correlation with R brand NT-proBNP reagent R2> 0.96.
Correlation between the low-value part (≤800pg/mL) and R brand NT-proBNP reagent R2> 0.97.
3.2 Fluorescence Immunochromatography (Lanthanide Eu Chelate Filling) Platform:
Clinical correlation between the high-value part and R brand NT-proBNP reagent R2 > 0.94.
Correlation between the low-value part (≤2500pg/ml) and R brand NT-proBNP reagent is R2 > 0.94.
4 Test Plan
4.1.1 Chemiluminescence (Acridinium Ester) Platform
4.1.1 Combination 1
- Antigen to be tested: NT-proBNP antigen (Cat. No.: [EKY0089E]), sample/standard adding volume: 20ul.
- Reagent 1: N-terminal brain natriuretic peptide precursor (NT-proBNP) coating (Cat. No.: [EKY0090L-1mg]) - magnetic bead dilution, dosage 50ul/test.
- Reagent 2: N-terminal brain natriuretic peptide precursor (NT-proBNP) label (Cat. No.: [EKY0090M-1mg]) - acridinium ester dilution, dosage 50ul/test.
- Reaction conditions: incubation temperature 37°C, sample + reagent 1 + reagent 2 (10 min).
- Luminescence instrument used in the test: Cosmai SMART 500S; chemiluminescence instrument applicable for various acridinium ester systems (e.g., Sichuan Mike, Shenzhen Yahuilong, Shandong Kanghua Bio, Jidan Bio, Nanjing Norman, etc.).
4.1.2 Combination 2:
- Antigen to be tested: NT-proBNP antigen (Cat. No.: [EKY0089F]), sample/standard adding volume: 20ul.
- Reagent 1: N-terminal brain natriuretic peptide precursor (NT-proBNP) coating (Cat. No.: [EKY0090P-1mg]) - magnetic bead dilution, dosage 50ul/test.
- Reagent 2: N-terminal brain natriuretic peptide precursor (NT-proBNP) label (Cat. No.: [EKY0090Q-1mg]) - acridinium ester dilution, dosage 50ul/test.
- Reaction conditions: incubation temperature 37°C, sample + reagent 1 + reagent 2 (10 min).
- Luminescence instrument used in the test: Cosmai SMART 500S; chemiluminescence instrument applicable for various acridinium ester systems (such as Sichuan Mike, Shenzhen Yahuilong, Shandong Kanghua Bio, Getein Bio, Nanjing Norman, etc.)
4.2 Fluorescence Immunochromatography (Lanthanide Eu Chelate Filling) Platform:
Antigen to be tested: NT-proBNP antigen (Cat. No.: [EKY0089E/EKY0089F]), sample volume: 10ul.
Reaction conditions: mix sample + fluorescent solution, load sample, reaction time (10min-15min).
Fluorescence instrument used in the test: Hemai FIC-S100; applicable to various fluorescence systems (excitation wavelength 365nm, emission wavelength 615nm) (such as Guangzhou Labsim, etc.)
5 Product information
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Product name
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Item number
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Recommended pairing
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N-terminal pro-brain natriuretic peptide (NT-proBNP) coating
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EKY0090L-1mg
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N-terminal pro-brain natriuretic peptide (NT-proBNP) coated (Cat. No.: EKY0090L-1mg) coated carboxyl magnetic beads + N-terminal pro-brain natriuretic peptide (NT-proBNP) label (Cat. No.: EKY0090M-1mg) label Acridinium ester（Luminous Pairing Set1）
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N-terminal pro-brain natriuretic peptide (NT-proBNP) labeling
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EKY0090M-1mg
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N-terminal pro-brain natriuretic peptide (NT-proBNP) coating
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EKY0090P-1mg
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N-terminal pro-brain natriuretic peptide (NT-proBNP) coated (Cat. No.: EKY0090P-1mg) coated carboxyl magnetic beads + N-terminal pro-brain natriuretic peptide (NT-proBNP) label (Cat. No.: EKY0090Q-1mg) label Acridinium ester（Luminous Pairing Set2）
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N-terminal pro-brain natriuretic peptide (NT-proBNP) labeling
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EKY0090Q-1mg
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N-terminal pro-brain natriuretic peptide (NT-proBNP) coating
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DKY0090O-1mg
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N-terminal pro-brain natriuretic peptide (NT-proBNP) labeled (Cat. No.: DKY0090N-1mg) labeled fluorescent microsphere + N-terminal pro-brain natriuretic peptide (NT-proBNP) coated (Cat. No.: DKY0090O-1mg) scratched （Fluorescence chromatography pair combination）
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N-terminal pro-brain natriuretic peptide (NT-proBNP) labeling
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DKY0090N-1mg
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NT-proBNP antigen
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EKY0089E
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Calibrator or Quality Control
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NT-proBNP antigen
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EKY0089F
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Calibrator or Quality Control
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N-terminal pro-brain natriuretic peptide (NT-proBNP) coating (Cat. No.: EKY0090L-1mg) - Carboxy magnetic bead concentrate
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DKY1309A
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N-terminal pro-brain natriuretic peptide (NT-proBNP) label (Cat. No.: EKY0090M-1mg)-acridinate concentrate
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DKY1309B
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N-terminal pro-brain natriuretic peptide (NT-proBNP) coating (Cat. No.: EKY0090P-1mg) - Carboxy magnetic bead concentrate
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DKY1309D
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N-terminal pro-brain natriuretic peptide (NT-proBNP) label (Cat. No.: EKY0090Q-1mg) - acridinium ester concentrate
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DKY1309E
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N-terminal pro-brain natriuretic peptide (NT-proBNP) label (Cat. No.: DKY0090N-1mg) - Carboxy fluorescent microsphere concentrate
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DKY1309C
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Carboxy magnetic beads (superparamagnetic), 1um
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DEG0091B-1-100mg
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Carboxy magnetic beads (superparamagnetic), 3um
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DEG0091A-3-100mg
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Time-resolved fluorescent microspheres, carboxyl (200nm）
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AEG0786B
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For more product details, please contact us: service@seebio.cn or Phone: +86 21 58183719 or Wechat: +86 158 0195 7578