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Serum autoantibody detection kit

A technology for detection kits and autoantibodies, applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of low detection sensitivity and accuracy, and achieve the effect of improving accuracy and increasing sensitivity

Active Publication Date: 2014-06-18
HANGZHOU KAIBAOLUO BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This fully demonstrates that although the single detection of tumor markers has high specificity, the detection sensitivity and accuracy are low; while the combined detection of multiple antigens, although the specificity is relatively lower, the sensitivity and accuracy are significantly improved. improvement

Method used

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  • Serum autoantibody detection kit
  • Serum autoantibody detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 , Recombinant antigenic protein expression and purification

[0051] Using the human cDNA library (purchased from Invitrogen) as a template, PCR amplification was used to prepare the required recombinant antigen protein fragments, wherein the amino acid sequence information of the recombinant antigen protein is shown in Table 1, and the primer information for PCR amplification is as follows Table 2 shows.

[0052] Table 1. Amino acid sequence information of recombinant antigen protein

[0053] recombinant antigenic protein

Amino acid sequence number

p53

SEQ ID NO:1

Annexin1

SEQ ID NO:2

CAGE

SEQ ID NO:3

Cyclin D

SEQ ID NO:4

NY-ESO-1

SEQ ID NO:5

GBU4‐5

SEQ ID NO:6

HuD

SEQ ID NO:7

PGP9.5

SEQ ID NO:8

MDM2

SEQ ID NO:9

GAGE7

SEQ ID NO:10

XAGE1b

SEQ ID NO: 11

SOX2

SEQ ID NO:12

MAGE A1

SEQ ID NO: 13

MAGE A...

Embodiment 2

[0060] Example 2 1. Preparation of enzyme-linked immunosorbent assay kit for detecting autoantibodies

[0061] 2.1. The composition of the ELISA kit for detecting autoantibodies

[0062] The ELISA kit (antigen immobilized) for the detection of autoantibodies includes the following components:

[0063] 1. ELISA plates coated with different antigen protein combinations;

[0064] 2. Enzyme conjugate-horseradish peroxidase-labeled goat anti-human immunoglobulin, the concentration is 0.1 μg / ml;

[0065] 3. Positive quality control (standard product): human anti-c-Myc-labeled immunoglobulin G (purchased from Tribioscience);

[0066] 4. Negative quality control products;

[0067] 5. Serum dilution buffer;

[0068] 6. TMB developer;

[0069] 7. 20×antigen washing solution;

[0070] 8. Termination solution;

[0071] 9. Parafilm.

[0072] 2.2. ELISA plate preparation and ELISA detection of serum samples

[0073] The steps of enzyme plate preparation and ELISA detection are as...

Embodiment 3

[0090] Example 3 , Antigen combination and clinical significance

[0091] 3.1. Detection of the effectiveness of candidate antigenic proteins in Chinese lung cancer patients

[0092] The recombinantly expressed and purified antigenic protein in Example 1 was tested for its effectiveness in Chinese lung cancer patients according to the method described in Example 2, including p53, Annexin1, CAGE, NY-ESO-1, HuD, Cyclin D, PGP9 .5, MDM2, GAGE7, GBU4‐5, XAGE1b, SOX2, MAGE A1, P62 and MAGE A4.

[0093] The result is shown in Figure 2, where:

[0094] 1. The OD450 of autoantibodies corresponding to p53 in patients with lung cancer was significantly higher than that in normal people and benign lung diseases, and 2 cases were positive in patients with benign lung diseases;

[0095] 2. The OD450 of autoantibodies corresponding to Mdm2 in lung cancer patients is significantly higher than that of normal people and benign lung diseases, and some samples in patients with benign lung di...

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Abstract

The invention discloses a detection kit used for detecting the serum autoantibody of mammals. The detection kit comprises an antigen protein, wherein the antigen protein is a combination of five or more out of p53, Annexin1, CAGE, NY-ESO-1, HuD, Cyclin D, PGP9.5, GBU4-5, MDM2, GAGE7, XAGE1b, SOX2, MAGE A1 and MAGE A4. The detection kit adopts a group of novel antigen combination corresponding to the autoantibody of a biomarker related to cancers, utilizes biotins to form characteristics of a polymer to envelope the antigen protein, and uses an anti-human label peptide as a standard of quantitative detection, thus increasing the detection sensibility and accuracy of the serum autoantibody, and providing an optimized detection method for using the autoantibody to carry out cancer diagnosis.

Description

technical field [0001] The invention belongs to the field of antibody detection, and in particular relates to a new serum autoantibody detection kit. Background technique [0002] According to the "2012 China Cancer Registration Annual Report" issued by the China Cancer Registration Center, there are about 3.12 million new cancer cases each year, an average of 8,550 people per day, and 6 people diagnosed with cancer every minute. Lung cancer ranks first in the incidence of malignant tumors in the country, followed by gastric cancer, colorectal cancer, liver cancer and esophageal cancer. The top 10 malignant tumors account for 76.39% of all malignant tumors. Lung cancer still ranks first in the death of malignant tumors in the country, followed by liver cancer, gastric cancer, esophageal cancer and colorectal cancer. The top 10 malignant tumors account for 84.27% of all malignant tumors. At the same time, the types of cancer also show regional characteristics. The high incid...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/574G01N33/6893G01N33/96G01N2800/7028
Inventor 孙苏彭
Owner HANGZHOU KAIBAOLUO BIOLOGICAL SCI & TECH
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