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Reagent kit for detecting serum HER2 and application

Inactive Publication Date: 2015-12-16
ANHUI PROVINCIAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In view of the problem that the currently used HER2 serum detection kits are easily interfered by other substances in the serum, resulting in low accuracy and stability of the detection results, the present invention provides a new kit that can significantly improve this problem

Method used

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  • Reagent kit for detecting serum HER2 and application
  • Reagent kit for detecting serum HER2 and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030]After diluting trastuzumab to 2 μg / mL, add 50 μL to each well and spread the plate overnight at 4°C. After washing the plate three times, block it with 1% BSA at room temperature for 1 hour; Dilute the special sample diluent to 20ng / mL, and use seven tubes to make 2-fold dilutions to form a gradient solution containing ECD, and wash the plate (the buffer used for each plate wash contains 5 / 10,000 concentration of Tween20, pH7.0 Phosphate buffer solution, the same buffer solution used for washing the plate below) three times, then add the gradient solution, 50uL per well, shake for 1 hour, then wash the plate three times, add biotin-A18, 50uL per hole, shake for 1 hour After washing the plate three times, add 50 uL of horseradish peroxidase to each well and shake for 1 hour. After washing the plate three times, add 50 uL of TMB chromogenic solution (manufactured by ThermoFisher) to each well. After 4 minutes of color development in the dark, add sulfuric acid stop solution...

Embodiment 2

[0032] Dilute trastuzumab to 5 μg / mL, add 50 μL to each well, plate at 4 °C overnight, wash the plate three times, add 300 μL 1% BSA solution to each well, block at 37 °C for 1 hour, add 0.4 mg / mL ECD in 1% BSA+ Serum (10:1), serum-specific sample diluent + serum (10:1), serum-specific sample diluent + fetal bovine serum (10:1) was diluted to a solution of 20ng / mL, and then seven tubes were used for 2 Double dilution to obtain a gradient solution, wash the plate (the buffer used for each plate wash is a phosphate buffer containing 5 / 10,000 concentration Tween20, pH7.0, and the same buffer used for the following plate wash) three times and then add Gradient solution was shaken at 50 μL per well for 1 hour. After washing the plate, 50 μL of biotin-A18 was added to each well for shaking for 1 hour. After washing the plate, 50 μL of horseradish peroxidase was added to each well for shaking for 1 hour. After washing the plate three times, TMB ( That is, 3,3',5,5'-Tetramethylbenzidi...

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Abstract

The invention provides a reagent kit for detecting serum HER2 and application. The reagent kit comprises HER2-resistant monoclonal antibody trastuzumab and biotinylated A18 which are paired. The invention further provides application of HER2-resistant antibodies and special sample diluent for serum to detect serum HER2 and application of manufacturing of the reagent kit for detecting serum HER2. The reagent kit for detecting serum HER2 and application can serve as a non-invasive, simple, convenient and fast method for diagnosing and detecting HER2 positive tumor disease course development.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine. Specifically, the present invention relates to a test kit for detecting the content of epidermal growth factor receptor 2 (HER2) in the serum of tumor patients, especially breast cancer, and its application, for example, for the detection by enzyme-linked immunosorbent assay (ELISA) A kit for detecting the content of HER2 in serum of patients with breast cancer and its application. More specifically, the present invention relates to a kit used in an ELISA detection method that can quantitatively detect the amount of HER2 in the serum of tumor patients, especially breast cancer patients, and its application in the diagnosis and treatment of patients, especially breast cancer patients. use in . Background technique [0002] Tumor is one of the major diseases seriously affecting public health. At present, there are nearly 2.2 million new cancer patients in my country every year, and mor...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/577G01N2800/365
Inventor 沈国栋胡世莲刘兢程民王璐龚欢秀徐婷娟
Owner ANHUI PROVINCIAL HOSPITAL
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