Kit for detecting natural bispecific antibody resisting thyroid peroxidase and thyroglobulin

A thyroglobulin and bispecific antibody technology, applied in the field of clinical laboratory science, can solve the problems of unfavorable promotion, use, biohazard and the like, and achieve the effects of convenient use, simple operation and strong specificity

Inactive Publication Date: 2014-02-26
BEIJING HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This detection method has high sensitivity and specificity, but there are obvious deficiencies. The detection process uses radioactive elements, ...

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  • Kit for detecting natural bispecific antibody resisting thyroid peroxidase and thyroglobulin
  • Kit for detecting natural bispecific antibody resisting thyroid peroxidase and thyroglobulin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: A kit for detecting anti-TPO and Tg bispecific antibodies

[0054] 1. Kit composition

[0055] 1. Human thyroglobulin (human Tg protein) coated microwell plate;

[0056] 2. Horseradish peroxidase-labeled human thyroid peroxidase (HRP-labeled human TPO protein): Beijing Yiqiao Shenzhou Biotechnology Co., Ltd., China;

[0057] 3. Washing solution (PBST): PBS solution containing 0.05% Tween-20; (add 500 μL Tween-20 to 1L PBS, mix well)

[0058] 4. Diluent: PBS containing 20% ​​(v / v) calf serum;

[0059] 5. Substrate solution: 3′,3′,5′,5′,-tetramethylbenzidine (TMB);

[0060] 6. Positive control solution: Serum from patients with Hashimoto’s thyroiditis containing natural bispecific antibodies against thyroid peroxidase and thyroglobulin;

[0061] 7. Negative control solution: healthy human serum;

[0062] 8. Stop solution: 0.5M sulfuric acid.

[0063] 2. Conditions and methods for preparing human thyroglobulin-coated microwell plates:

[0064] A: Micropla...

Embodiment 2

[0070] Example 2: Detection of anti-TPO and Tg bispecific antibodies

[0071] 1. Materials

[0072] 1. the test kit described in embodiment 1

[0073] 2. Serum: 136 sera of Hashimoto’s thyroid patients, 92 disease control sera (including 23 cases of hyperthyroidism, 20 cases of hypothyroidism, 22 cases of thyroid nodules, 20 cases of goiter, and 7 cases of thyroid cancer), 99 cases of healthy human serum. Stored at -80°C until analysis.

[0074] 2. Method

[0075] 1. Detection of anti-TPO and Tg bispecific antibodies by double-antigen sandwich method

[0076] (1) All materials and reagents were equilibrated to room temperature before measurement.

[0077] (2) Draw 100 μL of serum with a sample gun and add it to the human Tg protein-coated microwell plate provided in the kit, make duplicate wells for each sample, and incubate at 37°C for 1 hour;

[0078] (3) Wash 3 times with washing solution (PBST) and pat dry.

[0079] (4) The HRP-labeled human TPO protein was diluted 1...

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Abstract

The invention relates to a kit for detecting a natural bispecific antibody resisting thyroid peroxidase (TPO) and thyroglobulin (Tg), and belongs to the field of clinical laboratory science. Through detection on a patient serum, a disease control serum and a healthy control serum, the invention finds that detection rate of the anti-TPO and anti-Tg bispecific antibody in the patients is significantly higher than the control group, thereby showing statistical significance. The invention has the advantages that the detection kit described in the invention provides a simple and practicable detection mode for bispecific antibody, and the kit can be applied to detection of all kinds of bispecific antibodies.

Description

technical field [0001] The invention relates to a kit for detecting natural bispecific antibodies against thyroid peroxidase and thyroglobulin, belonging to the field of clinical laboratory science. Background technique [0002] The well-known antibody (Antibody, Ab) is a tetrapeptide chain structure composed of two identical heavy chains and two identical light chains connected by interchain disulfide bonds. X-ray crystallography shows a "Y" shape structure. Each antibody molecule has two identical antigen binding units (Antigen binding fragment, Fab) composed of light chain variable region and heavy chain variable region. Bispecific antibody (Bispecific antibody, BsAb) is a brand-new antibody concept, which refers to an immunoglobulin molecule containing two different antigen-binding sites. The first proposal refers to artificially synthesized anti-tumor drugs, not natural exist. In 1999, Dutch scholar Schuurman et al. used radioimmunoassay methods to discover a drug th...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/68
CPCG01N33/573G01N33/6893G01N2800/046G01N2800/7095
Inventor 李金明李文丽
Owner BEIJING HOSPITAL
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