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Reagent for investigating serum beta 2-glycoprotein I and its autoantibody

A technology of autoantibodies and glycoproteins, applied in biological testing, material inspection products, microbial measurement/inspection, etc., can solve the problems of lack of effective drugs, difficult diagnosis, poor specificity, etc.

Inactive Publication Date: 2006-11-29
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

current anti-beta 2 -The detection method of GPI antibody uses pure cardiolipin as an antigen to detect anti-β 2 -GPI antibody IgG, IgM and IgA, but in the detection of serum autoantibodies in clinical patients, due to high sensitivity and relatively poor specificity, more importantly, it fails to provide specific antiphospholipid syndrome pathogenesis-related Therefore, in the current laboratory diagnosis of antiphospholipid syndrome, there are still diagnostic difficulties, lack of effective drugs, and the mortality rate of patients is as high as 75%.

Method used

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  • Reagent for investigating serum beta 2-glycoprotein I and its autoantibody
  • Reagent for investigating serum beta 2-glycoprotein I and its autoantibody
  • Reagent for investigating serum beta 2-glycoprotein I and its autoantibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] beta 2 - ELISA assay of GPI-dependent anticardiolipin antibodies:

[0077] The concentration of cardiolipin (CL) was 50 μg / ml, and the amount of 50 μl / well was added to a 96-well Immulon 1B (Dynex Technologies Inc., Chantilly) ELISA culture plate. After physical drying and adsorption, 1% BSA was blocked, and then β 2 -GPI (15μg / ml, 50μl / well) and anti-β 2 -GPI antibody (Cof-22 and WB-CAL-I), co-incubated at room temperature for 1 hour, and finally incubated with horseradish peroxidase HRP-labeled anti-human IgG antibody for 1 hour, and measured the absorbance at 490 nm after color development. Between the above steps, the microtiter plate was fully washed with 0.05% Tween 20 PBS.

[0078] anti-beta 2 - ELISA assay for GPI-7-ketocholesteryl-9-carboxynonanoate complex antibody:

[0079] The chemically synthesized oxLDL derivative (Syn-oxLig-1) was added at a concentration of 50 μg / ml to a 96-well Immulon 1B (Dynex Technologies Inc., Chantilly) ELISA culture plate in a...

Embodiment 2

[0082] Anti-β in patients with systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS) 2 - Comparison of assays for GPI-ligand complex antibodies and anticardiolipin antibodies

[0083] Cardiolipin (CL) or a chemically synthesized oxLDL derivative (Syn-oxLig-1) at a concentration of 50 μg / ml was added to a 96-well Immulon 1B (Dynex Technologies Inc., Chantilly) ELISA culture plate in an amount of 50 μl / well, and physically dried After adsorption, 1% BSA was blocked, followed by sequential addition of β 2 -GPI (15 μg / ml, 50 μl / well) and serum samples from patients with SLE and APS (diluted 1:100 with 0.3% PBS, 50 μl / well), co-incubated for 1 hour at room temperature, and finally mixed with horseradish peroxidase HRP-labeled anti-human IgG antibody was incubated for 1 hour, and the absorbance at 490 nm was measured after color development. Between the above steps, the microtiter plate was fully washed with 0.05% Tween 20 PBS.

[0084] The above experimental res...

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Abstract

The invention relates to separation or chemical synthesis of a low-density fat protein oxidizing derivative 7-ketocholesteryl-9-carboxynonanoate specially combining with serum beta2-glycoprotein I (beta2-GPI); and usage of the derivative as substrate to detect and separate and purify the beta2-glycoprotein I; and an antibody detecting reagent for determining antibodies related with increase or decrease of danger in arteriosclerosis and thrombus formation related with beta2-GPI.

Description

Technical field: [0001] The invention relates to an oxidized low-density lipoprotein derivative compound 7-ketocholesteryl-9-carboxynonanoate specifically combined with serum β2-glycoprotein I (β2-GPI); the derivative is used as a substrate It is used to detect and isolate and purify β2-glycoprotein I; to determine whether there are diseases related to β2-glycoprotein, especially to determine whether there are antibodies related to the increase or decrease in the risk of arteriosclerosis and thrombosis, thereby to detect diseases. technical background: [0002] Antiphospholipid syndrome (APS), which belongs to systemic lupus erythematosus, is a clinically refractory type of autoimmune disease, the main clinical symptoms of which are arteriosclerosis and the formation of arteriovenous thrombosis, habit Sexual abortion and thrombocytopenia and other symptoms. The serological feature is the presence of numerous autoantibodies. [0003] Among them, anticardiolipin antibodies ...

Claims

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Application Information

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IPC IPC(8): G01N33/53C12Q1/00
Inventor 刘庆平刘华
Owner DALIAN UNIV
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