Single-reagent heparin binding protein detection reagent kit and method for preparing same

A technology of heparin-binding protein and detection kit, which is applied in measurement device, scattering characteristic measurement, color/spectral characteristic measurement, etc., can solve the problems of detection reagent sensitivity, specificity, and unsatisfactory linear range, and achieve a wide measurement linear range. , Simple operation, the effect of simplifying the test program

Inactive Publication Date: 2018-05-18
SUZHOU KANGHESHUN MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Many problems make the detection reagent obtained by this invention unsatisfactory in terms of sensitivity, specifi

Method used

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  • Single-reagent heparin binding protein detection reagent kit and method for preparing same
  • Single-reagent heparin binding protein detection reagent kit and method for preparing same
  • Single-reagent heparin binding protein detection reagent kit and method for preparing same

Examples

Experimental program
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Effect test

Embodiment 1

[0045] 1) Preparation of latex reagent (300nm, reaction solution) labeled with combination of monoclonal antibody and polyclonal antibody

[0046] Polystyrene latex particles with a carboxyl group on the surface (classification number K030, manufacturer Merck Millipore) with a particle size of 300 nm were labeled by the intermediate ester method. Dilute the microparticles with a particle size of 300nm and carboxyl groups on the surface to 1% in MES buffer solution, and stir at room temperature. Add EDC and Sulfo-NHS solid powder, stir for 2 hours and then centrifuge at 10000RPM for 20 minutes. The latex was washed twice with MES buffer solution, resuspended and divided into two parts. One part was added with HBP monoclonal antibody, stirred and reacted for 2 hours, and the other part was added with HBP polyclonal antibody, stirred and reacted for 2 hours. Both latexes were then centrifuged at 10,000 RPM for 20 minutes, and the supernatant was discarded. The two sets of prec...

Embodiment 2

[0054] 1) Preparation of a pair of monoclonal antibody-labeled latex reagents (200nm, reaction solution)

[0055]200nm polystyrene latex particles with carboxyl groups on the surface were labeled by the intermediate ester method (classification number H20021C, manufacturer Holmes). Microparticles with a particle size of 200 nm were diluted to 1% in PBS buffer solution and stirred at room temperature. Add EDC and Sulfo-NHS solid powder, stir for 2 hours and then centrifuge at 10000RPM for 20 minutes. A pair of HBP monoclonal antibodies was added, and the reaction was stirred for 2 hours. The latex was then centrifuged at 10,000 RPM for 10 minutes, and the supernatant was discarded. The precipitated latex was resuspended in the stock solution and ultrasonically dispersed, stirred for 1 hour before use. In this example, the reaction solution includes latex particles, buffer solution, salt, stabilizer, suspending agent, preservative and surfactant, wherein the concentration of ...

Embodiment 3

[0059] 1) Preparation of polyclonal antibody-labeled latex reagent (120nm, reaction solution)

[0060] 120nm polystyrene latex particles (classification number PS19021, manufacturer Merck Millipore) were labeled by physical adsorption. Microparticles with a particle size of 120 nm were diluted to 1% in PBS buffer solution and stirred at room temperature. Add EDC and Sulfo-NHS solid powder, stir for 2 hours and then centrifuge at 10000RPM for 20 minutes. Add HBP polyclonal antibody, and stir for 2 hours. The latex was then centrifuged at 10,000 RPM for 10 minutes, and the supernatant was discarded. The precipitated latex was resuspended in the stock solution and ultrasonically dispersed, stirred for 1 hour before use. In this example, the reaction solution includes latex particles, buffer solution, salt, stabilizer, suspending agent and preservative, wherein the concentration of latex particles is 0.05%, the buffer solution is 100mM Hepes pH=7.8, and the stabilizer is 1% bov...

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Abstract

The invention discloses a single-reagent heparin binding protein (HBP) detection reagent kit and a method for preparing the same. The single-reagent heparin binding protein detection reagent kit is asingle-reagent detection reagent kit. Main components of the single-reagent detection reagent kit include reaction liquid, standard products with HBP and quality control products. The reaction liquidcomprises HBP antibody labeling latex particles and further comprises buffer solution, surfactants, salt, stabilizers, suspending agents and preservatives. The invention further discloses a method fordetecting the concentration of the HBP in blood samples by the aid of the single-reagent heparin binding protein detection reagent kit. The single-reagent heparin binding protein detection reagent kit and the methods have the advantage that the single-reagent heparin binding protein detection reagent kit comprises a single reagent, is easy to operate, high in sensitivity and wide in linear rangeand can be widely applied to various transmission or scattering analyzers including universal biochemical analyzers, specific protein analyzers and the like.

Description

technical field [0001] The invention belongs to the field of medical immunodiagnostic reagents, in particular to a detection reagent for heparin-binding protein and a preparation method thereof. Background technique [0002] Heparin Binding Protein (HBP for short) is a granular protein secreted by neutrophils, which has a bactericidal effect and is also involved in the regulation of inflammatory responses and blood coagulation processes. In 1984, Shafer et al. first discovered and isolated this protein, and named it CAP37 (Cationic Antimicrobial Protein, molecular weight 37k) according to its electropositive characteristics and bactericidal function. Later, azurocidin and heparin-binding protein (HBP-Heparin Binding Protein), which has a strong binding ability to heparin, were isolated from the azurophilic blue granules in polymorphonuclear leukocytes. Further studies on protein structure and gene sequencing show that these three proteins are actually the same protein, whic...

Claims

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Application Information

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IPC IPC(8): G01N21/82G01N21/31G01N21/51
CPCG01N21/82G01N21/31G01N21/51G01N2021/825
Inventor 陈胜胜刘向晖王明
Owner SUZHOU KANGHESHUN MEDICAL TECH
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