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Hyaluronic acid chemiluminescence immunoassay kit and preparation method thereof

A detection kit, the technology of uric acid chemistry, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc., can solve the problems of low performance such as sensitivity and linear range, and reduce manual operation. The effect of error, large specific surface area and small systematic error

Inactive Publication Date: 2021-01-12
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main technical principle of these two detection methods is the double-antibody sandwich method, and the performance such as sensitivity and linear range is low.

Method used

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  • Hyaluronic acid chemiluminescence immunoassay kit and preparation method thereof
  • Hyaluronic acid chemiluminescence immunoassay kit and preparation method thereof
  • Hyaluronic acid chemiluminescence immunoassay kit and preparation method thereof

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preparation example Construction

[0055] figure 1 It is a design diagram of the preparation process of the hyaluronic acid chemiluminescent immunoassay kit of the present invention, including: screening of antibody raw materials: preparation of chemiluminescence labeling and reagent buffer; magnetic particle and antigen coating, preparation of solid-phase reagents; antigen raw materials Screening, preparation of calibrators, final assembly of reagents and supporting calibrators. The preparation method of the hyaluronic acid chemiluminescence immunoassay kit provided by the present invention specifically comprises the following steps:

[0056] Step 1, preparation of R1 reagent:

[0057] After the replacement of the hyaluronic acid antigen, suspend the magnetic particles and the hyaluronic acid antigen overnight, add a blocking agent, collect the magnetic field, and take the supernatant to obtain 1% to 3% coated magnetic particles of the hyaluronic acid antigen; 100-500mM Bistris, 0.05%-0.1% Tween-20, 0.05%-0....

Embodiment 1

[0064] Embodiment 1: the preparation of hyaluronic acid chemiluminescent immunoassay kit:

[0065] Preparation of R1 reagent: After replacing the hyaluronic acid antigen, suspend the Tosyl magnetic beads and the hyaluronic acid antigen overnight, add a blocking agent, collect the supernatant, and obtain the Tosyl magnetic beads coated with 2% hyaluronic acid antigen. particles. In 100mM Bistris, 0.05% Tween-20, 0.05% Proclin300, pH 6.0 buffer, the R1 reagent with a magnetic bead concentration of 0.072% was prepared.

[0066] Preparation of R2 reagent: put 500 μg hyaluronic acid-binding protein monoclonal antibody into a centrifuge tube, make sure that the antibody is at the bottom of the centrifuge tube, add carbonic acid buffer solution, mix well, add acridinium ester DMF solution after mixing, follow The labeling molar ratio of hyaluronan-binding protein monoclonal antibody to chemiluminescent marker is 1:10 for labeling, and the hyaluronan-binding protein chemiluminescent ...

Embodiment 2

[0068] Embodiment 2: the preparation of hyaluronic acid chemiluminescent immunoassay kit:

[0069] Preparation of R1 reagent: After replacing the hyaluronic acid antigen, suspend the Tosyl magnetic beads and the hyaluronic acid antigen overnight, add a blocking agent, collect the supernatant, and obtain the Tosyl magnetic beads coated with 3% hyaluronic acid antigen. particles. In 500mM Bistris, 0.1% Tween-20, 0.1% Proclin300, pH 6.5 buffer, the R1 reagent with a magnetic bead concentration of 0.05% was prepared.

[0070] Preparation of R2 reagent: put 500 μg hyaluronic acid-binding protein monoclonal antibody into a centrifuge tube, make sure that the antibody is at the bottom of the centrifuge tube, add carbonic acid buffer solution, mix well, add acridinium ester DMF solution after mixing, follow The labeling molar ratio of hyaluronan-binding protein monoclonal antibody to chemiluminescent marker is 1:3 for labeling, and the hyaluronan-binding protein chemiluminescent mark...

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Abstract

The invention relates to a hyaluronic acid chemiluminescence immunoassay kit and a preparation method thereof, and belongs to the technical field of in-vitro detection. The detection kit comprises: abuffer solution of a hyaluronic acid antigen coated with R1 reagent magnetic particles; R2 is a buffer solution of a hyaluronic acid binding protein monoclonal antibody marked by a chemiluminescent marker; and R3 is a buffer solution of hyaluronic acid binding protein. The R1, R2 and R3 reagents are assembled into a finished product reagent, namely the hyaluronic acid chemiluminescence immunoassaykit, and the chemiluminescence immunoassay kit can take a full-automatic chemiluminescence immunoassay instrument as a detection tool to finish detection of hyaluronic acid by using a competition method. The chemiluminiscence immunoassay kit is matched with an instrument, so that the time required by clinical detection is shortened, and the detection sensitivity is improved.

Description

technical field [0001] The invention belongs to the technical field of in vitro detection, and in particular relates to a hyaluronic acid chemiluminescent immunoassay kit and a preparation method thereof. Background technique [0002] Hyaluronic acid (HA), also known as hyaluronic acid and hyaluronic acid, is the only sulfur-free mucopolysaccharide known so far. HA is synthesized and secreted by interstitial cells, and most of its metabolism is completed by liver endothelial cells. Chronic hepatitis is caused by liver metabolism. The capacity decreases, the clearance of hepatic sinusoidal endothelial cells decreases, and HA is released into the blood. In liver cirrhosis, the ability of endothelial cells to absorb and decompose HA is reduced, which increases the level of serum HA. Therefore, the serum HA level is a reference index to measure the degree of liver fibrosis activity. [0003] At present, the main methods for the determination of hyaluronic acid are enzyme-linke...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543G01N33/532G01N21/76
CPCG01N21/76G01N33/532G01N33/54326G01N33/577
Inventor 张旭东李冬梅高智玲李磊孟令敏孙成艳高威何浩会
Owner DIRUI MEDICAL TECH CO LTD
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