Reagent or kit for magnetic particle chemiluminescence detection of salivary liquefied carbohydrate chain antigen and application of reagent or kit

A technology of chemiluminescence detection and sugar chain antigen, which is applied in chemiluminescence/bioluminescence, analysis through chemical reaction of materials, biological testing, etc., can solve the problems of low detection sensitivity, HOOK effect, short reaction time, etc., and achieve High luminous efficiency, good repeatability and high accuracy

Pending Publication Date: 2021-06-11
HUNAN YONGHE YANGGUANG SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Immunoturbidimetry has defects such as low detection sensitivity and high-concentration samples are prone to HOOK effects. Although the enzymatic chemiluminescence method solves the sensitivity problem, it has a long reaction time. It is necessary to develop a rapid test method with a short reaction time to determine KL -6

Method used

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  • Reagent or kit for magnetic particle chemiluminescence detection of salivary liquefied carbohydrate chain antigen and application of reagent or kit
  • Reagent or kit for magnetic particle chemiluminescence detection of salivary liquefied carbohydrate chain antigen and application of reagent or kit
  • Reagent or kit for magnetic particle chemiluminescence detection of salivary liquefied carbohydrate chain antigen and application of reagent or kit

Examples

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preparation example Construction

[0034] In the present invention, the preparation method of the magnetic particle-labeled KL-6 antibody 1 preferably includes the following steps:

[0035] 1) After washing the magnetic beads with a magnetic bead cleaning solution, activate them with EDC and NHS to obtain activated magnetic beads;

[0036]2) Shake the activated magnetic beads with KL-6 antibody 1, wash, block, and remove the supernatant to obtain magnetic particle-labeled KL-6 antibody 1.

[0037] In the present invention, the magnetic bead cleaning solution preferably includes an aqueous solution of 2-(N-morpholino)ethanesulfonic acid containing volume concentration 0.01%~0.1% Proclin300, 0.2M, more preferably containing volume concentration 0.02 % Proclin 300, 0.2M aqueous solution of 2-(N-morpholino)ethanesulfonic acid. The volume ratio of the magnetic beads, EDC and NHS is preferably 1-3:1-2:1-2, more preferably 2:1.5:1.5. The solution concentration of EDC is preferably 50 mg / ml. The solution concentrati...

Embodiment 1

[0055] 1. Preparation of the basic solution of the salivary sugar chain antigen (KL-6) assay kit (magnetic particle chemiluminescence method)

[0056] 1.1 Preparation of magnetic bead preservation solution: 0.01M phosphate buffer (pH=7.4) 80%, calf serum 10%, glycerin 10%, Proclin 3000.1%, Tween 200.1%;

[0057] 1.2 Preparation of acridinium ester preservation solution: 0.01M phosphate buffer (pH=7.4) 70%, glycerol 30%, BSA 1%.

[0058] 2. Preparation of magnetic bead-coated antibody in sialoligated sugar chain antigen (KL-6) assay kit (magnetic particle chemiluminescence method)

[0059] 2.1 Add 200 μL of magnetic beads into a 2 mL centrifuge tube, magnetically separate for 3 minutes, and then remove the supernatant.

[0060] 2.2 Add 400 μL of magnetic bead washing solution to the centrifuge tube, vortex to mix, remove the supernatant by magnetic separation, and wash twice.

[0061] 2.3 Weigh a certain amount of EDC, add a certain volume of MES buffer to make a 50mg / ml solu...

Embodiment 2

[0077] The using method of the kit prepared in embodiment 1 is as follows

[0078] 1. Take the reagent out of the box, remove the cap of the reagent bottle, cover the latex cap, and place it in the corresponding position of the setting;

[0079] 2. After the KL-6 calibrator is reconstituted, the reagent is calibrated.

[0080]3. After the calibration is completed, place the sample on the sample rack, and the instrument will automatically perform the test in the following order: 50 μL of magnetic particle reagent, 5 μL of sample, 50 μL of acridinium ester reagent, and read the corresponding concentration after incubation for 10 minutes.

[0081] 4. This kit confirms the key raw materials

[0082] The KL-6 antibodies provided by Beijing Boermai and Holmes were respectively coated with magnetic beads and labeled with acridinium ester, and prepared into different component reagents. The performance was evaluated by the lowest detection limit, and the best pairing was selected. A...

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Abstract

The invention provides a reagent or a kit for magnetic particle chemiluminescence detection of salivary liquefied carbohydrate chain antigen and application of the reagent or the kit, and belongs to the technical field of in-vitro diagnostic reagents. The reagent comprises a magnetic particle reagent and an acridinium ester reagent, wherein the magnetic particle reagent is composed of 2-8 [mu] g/ml of a magnetic particle labeled KL-6 antibody 1, 20%-30% of newborn bovine serum, 0.01%-0.1% of a preservative, 20%-30% of glycerin, 0.01%-0.1% of Tween 20, 0.1%-1% of BSA and 50%-70% of a PBS buffer solution, and the acridinium ester reagent is composed of 6-10 [mu] g/ml of an acridinium ester labeled KL-6 antibody 2, 10%-40% of glycerin, 0.5%-5% of BSA and 60%-90% of a PBS buffer solution. The KL-6 detection kit has the characteristics of high sensitivity, high precision and high accuracy, has excellent anti-interference capability, and does not have an obvious HOOK effect under a high-concentration sample of 40000IU/mL.

Description

technical field [0001] The invention belongs to the technical field of in vitro diagnostic reagents, and in particular relates to a reagent or kit for chemiluminescent detection of sialoglycosylated sugar chain antigen magnetic particles and an application thereof. Background technique [0002] Kohno discovered in 1985 that multiple strains of monoclonal antibodies were prepared by immunizing mice with a human lung adenocarcinoma cell line (VMRC-LCR), and the sialoligated sugar chain antigen recognized by the No. 6 antibody was named KL-6. KL-6 belongs to the epithelial mucin 1 (MUC1) classified as Cluster9. It is expressed on the surface of type II alveolar epithelial cells. It is only expressed in a small amount in normal lung tissue and terminal bronchiole epithelial cells. Enhanced cell expression. [0003] KL-6 is specific for judging the function of type Ⅱ alveolar epithelial cells. If the basement membrane of the lung is damaged, it can increase the permeability of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/68G01N21/76
CPCG01N21/76G01N33/54326G01N33/6893G01N2800/12
Inventor 刘健沈林陈汝彬靳启航
Owner HUNAN YONGHE YANGGUANG SCI & TECH
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