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Kit for simultaneously detecting gynecological tumor markers HER-2 and HE4

A technology for HER-2 and gynecological tumors, applied in measuring devices, scientific instruments, instruments, etc., can solve the problems of low sensitivity and specificity of kits, time-consuming human gynecological tumors, and low reliability of test results. Achieve the effects of screening and diagnosis, good antigenicity, and error reduction

Inactive Publication Date: 2019-02-15
JIANGSU INST OF NUCLEAR MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned kits have the defect that they can only detect a single gynecological tumor marker. The former is only for the detection of breast cancer, and the latter is only for the detection of ovarian cancer and uterine cancer. As a result, the clinical diagnosis, treatment and prognosis of human gynecological tumors cost a lot of money. More time, less efficiency
At the same time, the sensitivity and specificity of the above-mentioned kits are not high, and multiple separate detections are prone to large errors, which in turn leads to low reliability of the test results.

Method used

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  • Kit for simultaneously detecting gynecological tumor markers HER-2 and HE4
  • Kit for simultaneously detecting gynecological tumor markers HER-2 and HE4
  • Kit for simultaneously detecting gynecological tumor markers HER-2 and HE4

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Preparation of HER-2-T antibody and HE4-T antibody

[0040] The amino acid sequences shown in SEQ ID NO: 1 and SEQ ID NO: 2 were used as the HER-2-T-1 antigen and the HER-2-T-2 antigen to immunize animals, thereby preparing specific HER-2- T-1 and HER-2-T-2 antibodies. In the same way, the amino acid sequences shown in SEQ ID NO: 3 and SEQ ID NO: 4 are used as the HE4-T-1 antigen and HE4-T-2 antigen to immunize animals, thereby preparing specific HE4-T-1 using the above antigens. And HE4-T-2 antibody. HER-2-T-1 and HER-2-T-2 are abbreviated as HER-2-T-1 / 2, and HE4-T-1 and HE4-T-2 are abbreviated as HE4-T-1 / 2.

[0041] 1. Immunize animals to prepare monoclonal antibodies:

[0042] 1.1. Take the above-mentioned HER-2-T-1 antigen, HER-2-T-2 antigen, HE4-T-1 antigen and HE4-T-2 antigen (immunogen) respectively with an equal volume of Freund's complete adjuvant (Purchased from Sigma) After mixing thoroughly, immunize Balb / c mice with 50 μg antigen / mouse, and inject th...

Embodiment 2

[0052] Example 2: Specific identification of human HER-2-T-1 / 2 antibody and HE4-T-1 / 2 antibody

[0053] Detected by ELISA. Human HER-2, human HE4, carcinoembryonic antigen (CEA), neuron-specific enolase (NSE) (all purchased from Sigma) were used as the detection antigen to coat the ELISA plate, and then the prepared HER- The 2-T-1 / 2 monoclonal antibody and HE4-T-1 / 2 monoclonal antibody react specifically with the antigen plate. Normal BALB / c mouse serum is used as a negative control, and PBS solution is used as a blank control.

[0054] Results: The HER-2-T-1 / 2 monoclonal antibody only reacts positively with HER-2 (P / N> 2.1), HE4-T-1 / 2 monoclonal antibody only reacts positively with HE4 (P / N> 2.1), and the reaction with CEA and NSE is negative, indicating that the HER-2-T-1 / 2 monoclonal antibody and HE4-T-1 / 2 monoclonal antibody of the present invention have specificity.

[0055] The above method was used to identify the specificity of HER-2-T-1 / 2 polyclonal antibody and HE4-T-1 / 2 p...

Embodiment 3

[0056] Example 3 Preparation of a kit for simultaneous detection of gynecological tumor markers HER-2 and HE4

[0057] This embodiment provides a kit for simultaneously detecting the gynecological tumor markers HER-2 and HE4, which includes individually packaged: a lanthanide-labeled HER-2-T-1 antibody and a lanthanide-labeled HE4-T- 1 Antibody reagents, antibody-coated plates coated with HER-2-T-2 antibody and HE4-T-2 antibody, calibrator solution prepared with a series of concentrations of HER-2 and HE4 antigen, reaction buffer, concentrated Washing liquid and / or enhancing liquid.

[0058] Preparation of lanthanide-labeled HER-2-T-1 antibody solution (Eu-HER-2-T): Add 0.2 mg of europium labeling reagent (Eu-HER-2-T) to 1 mg of HER-2-T-1 monoclonal antibody 3+ -N 2 -[P-Isocyanic acid-benzyl]-diethylenetriaminetetraacetate sodium), mixed and magnetically stirred at 30°C for 20 hours. The next day, the mixture was subjected to cross-linked agarose gel 6B column chromatography, the ...

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Abstract

The invention belongs to the technical field of immunoassay, and particularly relates to a kit for simultaneously detecting gynecological tumor markers HER-2 and HE4 and use of the kit. The kit comprises a tracer-labeled HER-2 antibody and a tracer-labeled HE4 antibody. The adopted tracer-labeled HER-2 antibody and tracer-labeled HE4 antibody can be combined with a gynecological tumor marker HER-2or HE4 at high specificity, so that the kit can be utilized to simultaneously detect two tumor markers human HER-2 and HE4 by a double labelling time-resolved fluorescence immunoassay method, and helps simultaneous analysis of breast cancer, ovarian cancer and endometrial cancer, thereby improving efficiency of screening and clinical interpretation. Two results can be obtained based on a single operation of the kit, thereby improving test efficiency, reducing errors caused by individual test, and benefiting screening and diagnosis of gynecological tumors.

Description

Technical field [0001] The invention belongs to the technical field of immunoassays, and specifically relates to a kit for simultaneously detecting gynecological tumor markers HER-2 and HE4 and uses. Background technique [0002] Cancer has become a major issue threatening public health. In recent years, compared with men, the incidence of tumors in Chinese women has shown a significant annual increase of 2.2%. Among them, the incidence and mortality of female-specific breast cancer, ovarian cancer and uterine cancer are increasing year by year. Breast cancer accounts for 15% of new cancers in women, uterine cancer accounts for 3.6%, and ovarian cancer accounts for 3%. The rural areas have risen significantly. In order to cope with the high detection symptoms of Chinese women with idiopathic breast cancer, ovarian cancer and uterine cancer, early detection, early diagnosis and early treatment are one of the effective means to improve the survival rate of patients. Serological ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/533
CPCG01N33/533G01N33/57484
Inventor 张艺范俊周彬张珏韩婷婷谢敏浩
Owner JIANGSU INST OF NUCLEAR MEDICINE