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Anti-SS-B antibody IgG chemiluminescent immunoassay kit and preparation method thereof

A technology of chemiluminescent immunoassay and detection kit, which is applied in the field of in vitro diagnostic immunoassay, which can solve the problems of narrow detection linear range, low detection sensitivity, and high detection cost, and achieve the effect of wide linear range, high detection sensitivity, and easy operation

Inactive Publication Date: 2017-04-26
SHENZHEN YHLO BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The current anti-SS-B antibody IgG detection technology has the following disadvantages: high detection cost, low detection sensitivity, narrow detection linear range, low reproducibility, inability to quantify, complicated operation, etc.

Method used

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  • Anti-SS-B antibody IgG chemiluminescent immunoassay kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Preparation method of anti-SS-B antibody IgG chemiluminescent immunoassay kit

[0022] (1) Preparation of nano-magnetic beads coated with SS-B antigen:

[0023] Take 50mg of carboxylated magnetic particles (particle size: 0.05-1um) suspension, magnetically separate to remove the supernatant, resuspend with 0.02 M, pH 5.5MES buffer, add 0.5-2mL of newly prepared 10 mg / mL EDC Aqueous solution, activate the carboxyl groups on the surface of magnetic beads, add 3-5mg SS-B antigen, suspend at room temperature for 2-10 hours, magnetically separate, remove the supernatant, and resuspend with 0.1 M Tris buffer at pH 8.0 containing 2% BSA To 1mg / mL, SS-B antigen-coated magnetic particles were obtained, and each bottle was divided into 5mL and stored at 4°C for later use.

[0024] (2) Preparation of acridinium ester labeled with mouse anti-human IgG monoclonal antibody:

[0025] Take 50 uL 25mg / mL mouse anti-human IgG monoclonal antibody, add 150 uL 0.1-0.2 M carbona...

Embodiment 2

[0032] Embodiment 2: anti-SS-B antibody IgG chemiluminescent immunoassay method:

[0033] The present invention uses a fully automatic chemiluminescence immunoassay analyzer as a detection tool, and the methodology mode of the present invention is an indirect method, that is, the instrument sequentially adds 5 uL of sample, 95 uL of sample diluent, and 50 uL of SS-B antigen-coated magnetic particles After 10 min of reaction, magnetic separation was performed. Add 100 uL of acridinium ester coated with mouse anti-human IgG monoclonal antibody, react for 10 min, and perform magnetic separation. Luminescence reaction, and finally record the luminescence intensity, and calculate the anti-SS-B antibody IgG content of the tested sample from the standard curve.

Embodiment 3

[0034] Example 3: Performance evaluation of anti-SS-B antibody IgG chemiluminescence immunoassay kit

[0035] The detection curve is attached figure 1 .

[0036] Sensitivity detection:

[0037] The sensitivity of the anti-SS-B antibody IgG chemiluminescence immunochromatography kit was calculated according to the recommended experimental scheme in the CLSI EP17-A document, and the obtained sensitivity was 1ng / mL.

[0038] Linear detection:

[0039] Perform linear analysis on standard substances with concentrations of 5 AU / mL, 20AU / mL, 50 AU / mL, 90 AU / mL, and 165 AU / mL, and calculate the linear correlation coefficient, r=0.9996. In addition, the kit is resistant to SS-B The linear range of antibody IgG sample detection is 3-165AU / mL.

[0040] Precision determination:

[0041] Two anti-SS-B antibody IgG samples with a concentration of 2 AUmL and 100AU / mL were taken, and three parallel samples were made for each concentration of each sample, and three batches of kits were us...

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Abstract

The invention discloses an anti-SS-B antibody IgG chemiluminescent immunoassay kit, which comprises SS-B antigen coated magnetic particles, mouse-anti-human IgG monoclonal antibody coated acridinium ester, an anti-SS-B antibody IgG calibrated product, a pre-activating liquid, and an activating liquid. The invention further discloses a preparation method of the anti-SS-B antibody IgG chemiluminescent immunoassay kit. Compared with the conventional kits, the provided kit has the advantages of simple operation, high sensitivity, and wide detection range.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic immunoassay, and specifically, the invention provides a chemiluminescence immunoassay kit for anti-SS-B antibody IgG and a preparation method thereof. Background technique [0002] Single-strand binding protein (SSB, single-stranded binding protein) plays a very important role in DNA replication, recombination and repair. Its function is to stabilize the unwound single strand of DNA, prevent renaturation and protect the single-strand part from nucleases hydrolysis. It is a phosphorylated protein consisting of 408 amino acid residues with a molecular weight of 47 kDa. [0003] Anti-SSB antibodies are often detected in patients with Sjogren's syndrome and systemic lupus erythematosus, especially those with extraductal lesions (affecting the kidneys, lungs, and liver). It can also be detected years before the onset of clinical symptoms of lupus, and can also be detected in the serum of mothers of...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N33/532
CPCG01N33/532G01N33/54326G01N33/577G01N33/6842G01N2800/101G01N2800/104
Inventor 唐曙明陈海霞张昭李爽黄湘东马晓雯
Owner SHENZHEN YHLO BIOTECH
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