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Beta2-MG chemiluminescence immunoassay kit and preparation method therefor

A chemiluminescent immunoassay and reagent kit technology, applied in the field of immunoassay, can solve the problems affecting the detection of results, limited coating density, long reaction time, etc., and achieve the effect of short detection time, reliable detection means and high sensitivity

Inactive Publication Date: 2019-01-18
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above methods for quantitative detection of β2-MG have the following defects: 1) ELISA method adopts microporous plate as immobilization, limited coating density, low sensitivity, and long reaction time; 2) direct labeling antibody with isoluminol derivatives , using FITC antibody-FITC to indirectly couple FER antibody to the surface of magnetic beads, introducing more influencing factors and affecting the detection of results

Method used

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  • Beta2-MG chemiluminescence immunoassay kit and preparation method therefor
  • Beta2-MG chemiluminescence immunoassay kit and preparation method therefor
  • Beta2-MG chemiluminescence immunoassay kit and preparation method therefor

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Experimental program
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Effect test

Embodiment 1

[0050] Embodiment 1 main calibrator preparation

[0051] Dilute β2-MG with 100mM PBS buffer solution containing 20% ​​calf serum to form a calibrator, and aliquot it into concentrations of 0mg / L, 0.05mg / L, 0.2mg / L, 0.8mg / L, 2mg / L, respectively. L, 6 vials of 4.1 mg / L master calibrator. Based on the fitting curve of the main calibrator, the two-point calibration values ​​were obtained by back-value, and the concentrations of the two-point calibration of the enterprise were 0.1mg / L and 2mg / L respectively.

[0052] see figure 1 As shown, the dose-response calibration curve of the β2-MG chemiluminescence immunoassay assay kit has a linear correlation coefficient r≥0.9900, and the calibration is qualified. The specific parameters are listed in Table 1.

[0053] Table 1 Calibrator detection relative light value

[0054] Determination of calibrator concentration (mg / L)

Embodiment 2

[0055] Example 2 Preparation of β2-MG Chemiluminescence Immunoassay Assay Kit

[0056] Preparation of R1: Take 0.5 ml (50 mg) of streptavidin magnetic particle solution with a concentration of 100 mg / ml, add 10 ml of TBST solution and mix well for 10 minutes, then place on a magnetic separator until the supernatant is free of turbidity , discard the supernatant, and retain the magnetic particles. After repeated washing 3 times, the solid-phase reagent R1 with a magnetic bead concentration of 0.05% was prepared with 25mM Tris buffer containing 0.05% Tween and 0.02% sodium azide, pH 7.0, and stored at 2-8°C.

[0057] Preparation of R2: Put 500μg antibody into a centrifuge tube, make sure that the antibody is at the bottom of the centrifuge tube (centrifuge at room temperature for 20s), then add 50mM PBS buffer solution, mix well, after mixing, press antibody: acridine ester = 1 : 3 molar ratio, add the DMF solution of 2mg / mL acridinium ester, centrifuge 0.5 minute under room te...

Embodiment 3

[0059] Example 3 Preparation of β2-MG Chemiluminescence Immunoassay Assay Kit

[0060] Preparation of R1: Take 0.5 ml (50 mg) of streptavidin magnetic particle solution with a concentration of 100 mg / ml, add 10 ml of TBST solution and mix well for 10 minutes, then place on a magnetic separator until the supernatant is free of turbidity , discard the supernatant, and retain the magnetic particles. After repeated washing 3 times, the solid-phase reagent R1 with a magnetic bead concentration of 0.07% was prepared with 25mM Tris buffer containing 0.05% Tween and 0.02% sodium azide, pH 7.5, and stored at 2-8°C.

[0061] Preparation of R2: Put 500μg antibody into a centrifuge tube, make sure that the antibody is at the bottom of the centrifuge tube (centrifuge at room temperature for 20s), then add 100mM PBS buffer solution, mix well, after mixing, press antibody: acridinium ester = 1 : DMF solution of 10 molar ratio 2mg / mL acridinium ester, centrifugal 0.5 minute under room temper...

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Abstract

The invention discloses a beta2-MG chemiluminescence immunoassay kit and a preparation method therefor, belongs to the technical field of the immunoassay, and solves the problems of low technique sensitivity and high cost of the prior art. The kit comprises the following reagents: R1: the liquid containing streptavidin-coated magnetic beads; R2: the liquid containing the acridinium ester-labeled beta2-MG antibody; and R3: the liquid containing the biotin-labeled beta2-MG antibody. The beta2-MG chemiluminescence immunoassay kit of the invention has the advantages of high detection sensitivity,strong specificity, good repeatability, wide detection range and good stability, and can be widely used for clinical detection of the beta2-MG.

Description

technical field [0001] The invention relates to the technical field of immune analysis, in particular to a β2-MG chemiluminescence immunoassay assay kit and a preparation method thereof. Background technique [0002] β2-microglobulin (abbreviated as β2-MG) is a protein isolated from the urine of kidney patients by Berggard and Bearn in 1968. Because its molecular weight is 11800 and it appears in the β2 zone during electrophoresis, it is named β2-microglobulin. β2-MG is a single-chain polypeptide low-molecular protein composed of 100 amino acid residues produced by human nucleated cells. β2-MG is identical to the light chain of human leukocyte antigen (HLA), and contains a pair of disulfide bonds in the chain. β2-MG is non-covalently combined with the heavy chain of HLA-A, B, and C antigens and exists in the cell membrane superior. It is generally believed that except for mature erythrocytes and placental trophoblast cells, other cells contain β2-MG. Therefore, both norm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/553G01N33/58G01N33/532G01N21/76
CPCG01N33/6803G01N21/76G01N33/532G01N33/54326G01N33/553G01N33/58
Inventor 刘杨翟涛高威孙成艳何浩会
Owner DIRUI MEDICAL TECH CO LTD
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