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Method for detecting human immunoglobulin G4 and kit

A human immunoglobulin and kit technology, applied in the field of medical testing, can solve problems such as insufficient specificity of detection

Active Publication Date: 2019-10-01
山西瑞豪生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One possibility for a high basal value of human serum IgG4 is that the assay is not specific enough to include other immunoglobulins
Therefore, there is a potential risk for specific recognition of IgG4 changes in human serum

Method used

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  • Method for detecting human immunoglobulin G4 and kit
  • Method for detecting human immunoglobulin G4 and kit
  • Method for detecting human immunoglobulin G4 and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1: A kind of test kit that detects IgG4 antibody concentration

[0057] 1. Kit composition:

[0058] Table 1: Kit Composition

[0059]

[0060]

[0061] 2. Preparation method

[0062] 1. Preparation of ELISA plate

[0063] 1.1 Preparation of coating solution

[0064] Table 2

[0065]

[0066] *The IgG4Fc antibody in Table 2 is the first IgG4 monoclonal antibody, produced by Abcam, and the product numbers are ab1930.

[0067] 1.2 Coating of ELISA plate

[0068] Add 100 μL of coating solution to each well of the 96-well ELISA plate with a coating machine in the coating room of the 100,000-level area, and place it at 4°C overnight.

[0069] 1.3 Preparation of blocking solution

[0070] table 3

[0071]

[0072] 1.4 Sealing of the microtiter plate

[0073] Discard the coating solution and wash once (add 300uL 1×PBST to each well, let it stand for 2min, shake off the solution in the well, and pat dry). Add 200 μL of blocking solution to each ...

Embodiment 2

[0163] Example 2. Method for detecting human immunoglobulin G4 in a sample

[0164] 1. Materials and methods

[0165] 1. Materials:

[0166] (1) The test kit of embodiment 1

[0167] (2) The sample with known diluted IgG4 concentration is a patient sample, which comes from a male patient in the Department of Gastroenterology, First Hospital of Shanxi Medical University, whose IgG4 concentration was measured by this kit to be 0.401g / L.

[0168] (3) Patient samples to be tested: Serum samples were provided by the Gastroenterology Department of the First Hospital of Shanxi Medical University. Department of Gastroenterology, No. 1 Hospital, Tel: 0351-4639020.

[0169] 2. Method:

[0170] (1) Treat the sample to be tested: dilute the sample to be tested with a volume ratio of 1:10000, take 10 μL of the sample and add it to 1 mL of sample diluent, shake and mix, then take 10 μL of it and add it to 1 mL of sample diluent, shake and mix well ;

[0171] (2) Adding samples: Add di...

Embodiment 3

[0186] Embodiment 3. kit and method specific experiment

[0187] 1. Materials and methods

[0188] 1. Materials

[0189] (1) Example 1 kit

[0190] (2) IgG1, IgG2, IgG3 samples, IgG1 was purchased from Abcam, the product number is ab90283; IgG2 was purchased from Abcam, the product number is ab90284; IgG3 was purchased from Sino.Biological Inc, the product number is 13906-HNAH.

[0191] 2. Method

[0192] Dilute IgG1, IgG2, and IgG3 to the following concentrations of test samples: 1000ng / mL, 500ng / mL, 250ng / mL, 100ng / mL and 50ng / mL;

[0193]Use the kit of the present invention to measure IgG1, IgG2, IgG3 (1000ng / mL, 500ng / mL, 250ng / mL, 100ng / mL and 50ng / mL) with different concentrations respectively, and the method is the same as the detection method in Example 2.

[0194] 2. Results

[0195] The measurement results are shown in Table 9, and it was found that the OD values ​​were all close to the blank wells. From the results, it was confirmed that the method established...

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Abstract

The invention relates to a method for detecting human immunoglobulin G4 and a kit, and belongs to the technical field of medical inspection. According to the method for detecting the human immunoglobulin G4 (IgG4), two monoclonal antibodies with high-specificity anti-IgG4Fc fragments are used, wherein the first monoclonal antibody specific to IgG4 is combined with a solid-phase carrier while the second monoclonal antibody specific to IgG4 is marked by a peroxidase. By virtue of a bridging effect of IgG4 in a to-be-tested sample, a compound of first IgG4 monoclonal antibody-IgG4 enzyme marked second IgG4 monoclonal antibody is formed, and the compound is fixed on the solid-phase carrier. According to the method, the non-specific interfering substance components in the sample are removed byadopting a high-efficiency cleaning fluid. According to the method for detecting the content of the biological sample IgG4, the high stability and the high specificity of the quantitative detection ofthe IgG4 in the biological sample are realized, and accurately quantifying of the concentration of the to-be-detected object IgG4 in a human biological sample is realized.

Description

technical field [0001] The invention relates to a method and a kit for detecting human immunoglobulin G4, belonging to the technical field of medical testing. Background technique [0002] Human immunoglobulin G4 (referred to as human IgG4) is the fourth of the four subtypes of immunoglobulin G (IgG) in human blood. With its unique role in autoimmune diseases, it constitutes a variety of human autoimmune diseases. The basis of disease forms a new field in the clinical study of autoimmune diseases. IgG4-related autoimmune diseases are a type of diseases characterized by the production of IgG4 antibodies against specific antigens in the body. The pathogenesis of IgG4-related autoimmune diseases is significantly different from that of other immunoglobulins. Compared with other immunoglobulins, IgG4 has a sluggish immune response and has a monovalent functional structure. It slowly forms autoimmune antibodies after exposure to antigens, and the formed antibodies can compete wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/535G01N33/545
CPCG01N33/6854G01N33/535G01N33/545
Inventor 王敏兰牛林茹陈婷婷张旋旋奚瑞芳阎婧杨武
Owner 山西瑞豪生物科技有限公司
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