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NT-ProBNP detection kit and using method thereof

A detection kit and magnetic bead technology, which is applied in the field of NT-ProBNP detection kits, can solve problems such as narrow detection range and limited application, and achieve the effects of rapid diagnosis, improved detection speed, and expanded application range

Active Publication Date: 2018-02-02
NANTONG EGENS BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned NT-proBNP detection kit is conducive to the rapid detection of NT-proBNP in blood, and improves the detection sensitivity to a certain extent, but the detection range of the kit is only 0-20ng / ml, and the detection range is narrow, which limits its clinical application

Method used

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  • NT-ProBNP detection kit and using method thereof

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Embodiment 1

[0057] This embodiment provides a method for preparing an alkaline phosphatase-labeled NT-ProBNP antibody, comprising the following steps:

[0058] 1. Weigh an appropriate amount of Traut's reagent, and prepare a 1.376 mg / mL solution with pH 8.5, 100 mM triethanolamine buffer.

[0059] 2. Store the NT-ProBNP-labeled antibody (that is, the NT-ProBNP antibody) in a pH 8.5, 100mM triethanolamine buffer solution with an antibody concentration of 2mg / mL; add the Traut' prepared in step 1 to the above antibody solution s solution, the molar ratio of Traut's reagent to NT-ProBNP antibody was 30:1, mixed immediately, and left to react at 25°C for 15 minutes.

[0060] 3. Add 1M glycine solution to the antibody solution obtained after the activation reaction in step 2, and the molar ratio of glycine to Traut's reagent is 20:1; mix immediately, and stand at 25°C for 10 minutes for reaction.

[0061] 4. The NT-ProBNP-labeled antibody solution obtained after the reaction in step 3 was imm...

Embodiment 2

[0072] This embodiment provides a method for preparing an alkaline phosphatase-labeled NT-ProBNP antibody, comprising the following steps:

[0073] 1. Weigh an appropriate amount of Traut's reagent, and prepare a 1.376 mg / mL solution with pH 8.5, 100 mM triethanolamine buffer.

[0074] 2. Store the NT-ProBNP-labeled antibody (that is, the NT-ProBNP antibody) in pH 8.5, 100mM triethanolamine buffer, and the antibody concentration is 5mg / mL; add the Traut' prepared in step 1 to the above antibody solution s solution, the molar ratio of Traut's reagent to NT-ProBNP antibody was 15:1, mixed immediately, and left to react at 25°C for 15 minutes.

[0075] 3. Add 1M glycine solution to the antibody solution obtained after the activation reaction in step 2. The molar ratio of glycine to Traut's reagent added is 10:1; mix immediately, and stand at 25°C for 10 minutes for reaction.

[0076] 4. The NT-ProBNP-labeled antibody solution obtained after the reaction in step 3 was immediately...

Embodiment 3

[0087] This embodiment provides a method for preparing magnetic beads labeled with an NT-ProBNP antibody, comprising the following steps:

[0088] 1. Store the NT-ProBNP-coated antibody (that is, the NT-ProBNP antibody) in MES buffer at pH 5.0 and 100 mM, and the antibody concentration is 1 mg / mL;

[0089] 2. Take the carboxyl magnetic bead solution 100 times the weight of the antibody, the concentration of the carboxyl magnetic bead solution is 100 mg / ml, the particle size of the carboxyl magnetic bead is 3 μm, and the supernatant is discarded by magnetic separation;

[0090] 3. Washing: re-dissolve the magnetic beads obtained in the above steps with pH 5.0, 100 mM MES buffer, and discard the supernatant by magnetic separation.

[0091] 4. Repeat step 3 once.

[0092] 5. Add an appropriate amount of 100mM MES buffer to dissolve the magnetic beads obtained in step 4.

[0093] 6. Weigh an appropriate amount of NHS (N-hydroxysuccinimide) reagent and dissolve it with pH 5.0, 10...

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Abstract

The invention discloses an NT-ProBNP detection kit. The NT-ProBNP detection kit comprises a calibrator, a cleaning solution, a substrate solution, a pretreatment solution, an enzyme conjugate workingsolution and a magnetic bead conjugate working solution; the pretreatment solution contains pyridine, the enzyme conjugate working solution contains NT-ProBNP antibody labeled by enzyme, and the magnetic bead conjugate working solution contains magnetic beads labeled by the NT-ProBNP antibody. The NT-ProBNP detection kit can accurately measure NT-ProBNP in a whole blood sample, the lowest limit detection of the kit is 20 pg / ml, the linearity range is 20-5,000 pg / ml, the detection sensitivity is high, the linearity range is wide, and the result is accurate. The invention further discloses a using method of the NT-ProBNP detection kit. The NT-ProBNP detection kit is simple in using step, the detection time of an NT-ProBNP is shortened, and quick and sensitive detection of the NT-ProBNP is achieved.

Description

technical field [0001] The invention belongs to the technical field of immunochemical detection, and in particular relates to an NT-ProBNP detection kit and a use method thereof. Background technique [0002] B-type natriuetic peptide (BNP) is a cardiac hormone mainly synthesized and secreted by ventricular myocardium, BNP and atrial peptide (ANP), vascular endothelial cell secretion (CNP), renal tubular synthesis and secretion ( RNP) is a member of the natriuretic peptide family. BNP has the functions of diuresis, natriuresis, dilating blood vessels, inhibiting the renin-angiotensin-aldosterone system, inhibiting the release of ACTH, inhibiting the excessive response of the sympathetic nerve, and participating in the regulation of blood pressure, blood volume, and salt balance. Recent studies have shown that BNP also inhibits Myocardial fibrosis, inhibition of vascular smooth muscle cell proliferation and anti-coronary spasm. N-terminal B-type natriuretic peptide (NT-proB...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/325G01N2800/50
Inventor 王保君汤双双欧卫军苏玲玲
Owner NANTONG EGENS BIOTECH
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