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Kit for detecting human heparin binding protein and preparation method thereof

A heparin-binding protein and kit technology, applied in the field of detection of human heparin-binding protein, can solve the problems of not covering high-concentration HBP, low sensitivity, and only reaching 0.00-500.00ng/mL

Pending Publication Date: 2020-02-18
SHENZHEN AMTECH BIOENGINEERING LTD INC
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the latex-enhanced immunoturbidimetric method has problems such as low sensitivity and insufficiently wide detection linear range, such as patent applications CN107167589A and CN108152512A
In order to solve the problem of sensitivity and detection linear range, the patent application CN109613259A marked HBP monoclonal antibody and HBP polyclonal antibody on different latex microspheres respectively, hoping to improve the performance of latex-enhanced immunoturbidimetric method through the combination of HBP monoclonal antibody and polyclonal antibody. Sensitivity and detection linear range; however, the sensitivity in this patent application can only reach 1.00ng / mL, and the detection linear range can only reach 0.00-500.00ng / mL, which cannot cover high concentrations of HBP exceeding 500.00ng / mL

Method used

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  • Kit for detecting human heparin binding protein and preparation method thereof
  • Kit for detecting human heparin binding protein and preparation method thereof
  • Kit for detecting human heparin binding protein and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] 1. Materials and methods

[0062] 1. Main material

[0063] Tris (Tris), Bovine Serum Albumin (BSA), Sodium Azide, Macrogol 10K, Sodium Chloride, TritonX-100, 4-Hydroxyethylpiperazineethanesulfonic acid, Tween 20 , Tris-HCl, KCl, CaCl 2 , Mannitol are analytically pure or chemically pure reagents routinely used in laboratories. The latex microspheres were polystyrene microspheres with carboxyl groups purchased from Bangs Lab; in this example, two specifications of polystyrene microspheres with carboxyl groups were used for the test, that is, the particle size range was 80-180nm, and the surface Small spherical latex microspheres with a charge of 150-200μeq / g, and large spherical latex microspheres with a particle size range of 380-400nm and a surface charge of 20-50μeq / g. Human heparin-binding protein monoclonal antibody (abbreviated as HBP monoclonal antibody) was purchased from R&D Systems (USA). Human heparin-binding protein antigen (abbreviated as HBP antigen) w...

Embodiment 2

[0127] In this example, on the basis of Example 1, the particle size range of the large-sphere latex microspheres, the particle size range of the small-sphere latex microspheres, and the ratio of the two were tested. The results show that the particle size range of the large-sphere latex microspheres is 280-400nm, and the particle size range of the small-sphere latex microspheres is 80-180nm. The difference is at least 200nm, and the surface charge value of the small-sphere latex microsphere is at least 100μeq / g larger than that of the large-sphere latex microsphere, which can improve the detection sensitivity and linear range.

[0128] The particle size of large-sphere latex microspheres greater than 400nm is not conducive to the detection stability, while the particle size of less than 280nm is not conducive to the improvement of sensitivity; the particle size of small-sphere latex microspheres greater than 180nm or less than 80nm is not conducive to the improvement of the li...

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Abstract

The application discloses a kit for detecting human heparin binding protein and a preparation method thereof. The kit for latex enhanced immunoturbidimetry detection comprises a reagent 1, a reagent 2and a calibration product. The reagent 2 contains HBP-monoclonal-antibody-labeled large-sphere latex microspheres and HBP-monoclonal-antibody-labeled small-sphere latex microspheres, wherein a particle size difference between the large-sphere latex microsphere and the small-sphere latex microsphere is at least 200 nm and a surface charge difference between the small-sphere latex microspheres andthe large-sphere latex microspheres is at least 100 [mu] eq / g. According to the application, the large-sphere and small-sphere latex microspheres with the particle size difference value of at least 200nm and the surface charge difference value of at least 100 [mu]eq / g are used for HBP monoclonal antibody labeling to realize latex enhanced immunoturbidimetry detection, so that the detection sensitivity can be improved and the wider detection linear range can be obtained. A high-sensitivity and full-scale detection kit is provided for HBP clinical detection.

Description

technical field [0001] The present application relates to the field of detection of human heparin-binding protein, in particular to a kit for detection of human heparin-binding protein and a preparation method thereof. Background technique [0002] Human heparin-binding protein (HBP) is a granule protein derived from neutrophils, which is mainly released by polymorphonuclear neutrophils (PMN) by external stimuli. The content of HBP in normal human blood is very low, generally not exceeding 10ng / mL. When an infection occurs, some bacteria invade the blood vessels, and substances such as the bacteria themselves or the toxins released by the bacteria stimulate the neutrophils to release HBP, resulting in an increase in the HBP content in the blood. HBP can reach 20-30ng / mL in general infection. Severe infections in the ICU will be higher, can exceed 100ng / mL, and even reach 1000ng / mL. [0003] HBP can be used as a diagnostic indicator of local infection, an indicator of auxi...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/68G01N33/54313
Inventor 陈小茹吴向东
Owner SHENZHEN AMTECH BIOENGINEERING LTD INC
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