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Human heparin binding protein assay kit with high sensitivity and wide detection range

A heparin-binding protein and kit technology, applied in the field of immunodetection, can solve the problems of inaccurate quantification, long time consumption, insufficient sensitivity, etc.

Inactive Publication Date: 2019-04-12
BEIJING BEIER BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ELISA operation steps are cumbersome, time-consuming, and there are many human factors, which are easy to introduce errors; the chromatography reagent system has large variations, the quantification is not accurate enough, and the sensitivity is not enough; the latex-enhanced immunoturbidimetric reagent can be operated in a fully automatic mode on the biochemical analyzer , the system error is small, the precision is good, and the reagent stability is good, but the detection sensitivity of the existing reagents is not good enough, and the linear range is not wide enough

Method used

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  • Human heparin binding protein assay kit with high sensitivity and wide detection range
  • Human heparin binding protein assay kit with high sensitivity and wide detection range
  • Human heparin binding protein assay kit with high sensitivity and wide detection range

Examples

Experimental program
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Effect test

Embodiment 1

[0064] A human heparin-binding protein assay kit, comprising reagent 1, reagent 2 and human HBP calibrator with different concentration gradients; wherein,

[0065] Reagent 1 comprises the following components of final concentration: 20mM HEPES buffer solution (pH7.2), the BSA of 1.5% (w / v), the PEG6000 of 2% (w / v), the polyol of 5% (w / v) Polylysine, DTT of 10mM, KCl of 150mM, Tween-20 of 0.05% (w / v), Krovin300 of 0.1% (w / v), EDTA of 5mM;

[0066] Reagent 2 includes the components of the following final concentrations: carboxyl latex microspheres (average particle diameter is 273nm, microsphere final concentration is 0.015% (w / v)), 0.008% (w / v ) labeled human HBP monoclonal antibody, carboxyl latex microspheres labeled only with human HBP polyclonal antibody (the average particle diameter is 123nm, the final concentration of microspheres is 0.2% (w / v)), 0.045% (w / v) Labeled human HBP polyclonal antibody, 50mM HEPES buffer (pH7.4), 2% (w / v) BSA, 4% (w / v) sucrose, 1% (w / v) treh...

Embodiment 2

[0073] A human heparin-binding protein assay kit, comprising reagent 1, reagent 2 and human HBP calibrator with different concentration gradients; wherein,

[0074] Reagent 1 included the following final concentrations of components: 50 mM MOPS buffer (pH 7.4), 0.5% (w / v) BSA, 2.5% (w / v) PEG8000, 1.5% (w / v) Mw Dextran of 40000, TECP of 5mM, NaCl of 300mM, Tween-20 of 0.05% (w / v), ProClin300 of 0.1% (w / v), EDTA of 5mM;

[0075] Reagent 2 includes the components of the following final concentrations: carboxyl latex microspheres (average particle diameter is 223nm, microsphere final concentration is 0.02% (w / v)), 0.006% (w / v ) labeled human HBP monoclonal antibody, carboxyl latex microspheres labeled only with human HBP polyclonal antibody (average particle size is 88nm, final concentration of microspheres is 0.25% (w / v)), 0.04% (w / v) Labeled human HBP polyclonal antibody, 20mM PBS buffer (pH7.4), 0.5% (w / v) gelatin, 2% (w / v) sucrose, 3% (w / v) trehalose, 50mM chloride Potassium...

Embodiment 3

[0082] A human heparin-binding protein assay kit, comprising reagent 1, reagent 2 and human HBP calibrator with different concentration gradients; wherein,

[0083] Reagent 1 includes the following final concentrations of components: 20 mM PBS buffer (pH 7.2), 0.3% (w / v) BSA, 4% (w / v) PEG6000, 3% (w / v) Mw 10000 PVP, 20mM TECP, 50mM NH 4 Cl, 0.1% (w / v) Tween-80, 0.1% (w / v) ProClin300, 2mM EDTA;

[0084] Reagent 2 includes the components of the following final concentrations: carboxyl latex microspheres (average particle diameter is 309nm, microsphere final concentration is 0.01% (w / v)), 0.01% (w / v ) labeled human HBP monoclonal antibody, carboxyl latex microspheres labeled only with human HBP polyclonal antibody (average particle diameter is 83nm, final concentration of microspheres is 0.3% (w / v)), 0.035% (w / v) Labeled human HBP polyclonal antibody, 50mM TAPS buffer (pH8.0), 0.5% (w / v) gelatin, 0.5% (w / v) BSA, 2% (w / v) sucrose, 2% (w / v) Trehalose, 50 mM ammonium chloride, 0...

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Abstract

The embodiments of the invention relate to the field of immunoassay, and in particular to a human heparin binding protein assay kit with high sensitivity and wide detection range. The human heparin binding protein assay kit comprises: a reagent 1, a reagent 2 and human HBP calibrators with different concentration gradients. The reagent 2 comprises a carboxyl latex microsphere labeled only with human HBP monoclonal antibody and a carboxyl latex microsphere labeled only with human HBP polyclonal antibody. The average particle diameter of the carboxyl latex microsphere labeled only with the humanHBP monoclonal antibody > the average particle diameter of the carboxyl latex microsphere labeled only with the human HBP polyclonal antibody. The human heparin binding protein assay kit provided bythe invention can complete a single sample test within 10 minutes, and indexes of precision, accuracy and anti-interference are excellent. The invention can be used clinically to predict an organ dysfunction caused by sepsis, and can be used as an early diagnostic marker for sepsis, especially serious bacterial infection.

Description

technical field [0001] The invention relates to the field of immunoassay, in particular to a human heparin-binding protein assay kit with high sensitivity and wide detection range. Background technique [0002] Heparin-binding protein (HBP), also known as azurocidin or CAP37, is released by neutrophils after stimulation. It was first isolated and identified by Shafer et al. in 1984. There are 8 cysteine ​​residues in the 222 amino acids of HBP, and there are glycosylation sites on the 100th, 114th or 145th aspartic acid residue. [0003] The bactericidal ability of HBP and the ability to integrate with heparin are related to its strong positive charge. HBP is a member of the trypsin-like serine protease family that activates monocytes and macrophages and alters endothelial cell permeability, thereby inducing vascular leakage and tissue edema. HBP can predict organ dysfunction caused by sepsis, and can be used as an early diagnostic marker for sepsis, especially severe bact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6887
Inventor 不公告发明人
Owner BEIJING BEIER BIOENG
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