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A kit for detecting erbb2 protein and its preparation method

A technology of ERBB2 and kits, which is applied in the field of kits for detecting ERBB2 proteins and its preparation, can solve problems such as the ultra-trace detection of ERBB2 proteins, and achieve the effect of improving the luminescent signal

Active Publication Date: 2015-10-21
SUZHOU HYBIOME BIOMEDICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0022] The invention overcomes the technical problem that the current chemiluminescence method cannot meet the ultra-trace detection of ERBB2 protein, that is, combines the chemiluminescence technology with the biotin-avidin immunomagnification technology and the water-soluble 5,10,15-tris(4-pyridyl )-20-R-carboxyporphyrin is used as a marker to construct a chemiluminescent immunoassay to detect ERBB2 protein with high sensitivity, and a kit for detecting ERBB2 protein and its preparation method are provided

Method used

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  • A kit for detecting erbb2 protein and its preparation method
  • A kit for detecting erbb2 protein and its preparation method
  • A kit for detecting erbb2 protein and its preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0056] Example 1 Preparation of the kit for detecting ERBB2 protein of the present invention

[0057] 1. Preparation of calibrator matrix solution

[0058]

[0059] Weigh the above reagents into a clean container, add double distilled water to make up the volume, dissolve and mix well, and measure the pH value. Store at 2-8°C after labeling.

[0060] 2. Preparation of ERBB2 protein calibrator

[0061] Prepare calibrator with calibrator matrix solution and pure ERBB2 protein, the concentrations are 0, 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 100ng / mL, (wherein 0ng / mL is calibrator matrix solution, ERBB2 protein-free), prepare a series of concentration calibration products, each concentration calibration product is divided into 1mL / vials of 3mL size, freeze-dried, and stored at -20°C.

[0062] 3. Preparation of formula diluent

[0063]

[0064] 4. Preparation of streptavidin-coated solid-phase magnetic particles

[0065] (1) Weigh 250 mg of amino-terminal magnetic particle...

Embodiment 2

[0096] Example 2 Prepare the kit for detecting ERBB2 protein of the present invention

[0097] 4. Preparation of streptavidin-coated solid-phase plastic particles

[0098] (1) Weigh 250 mg of amino-terminal plastic particles and place them in a beaker;

[0099] (2) Add 5 mL each of methanol and acetone to wash repeatedly 5 times;

[0100] (3) centrifugal separation, discard the supernatant;

[0101] (4) Add 10 mL of 0.2% 1,4-phenylene diisothiocyanate (PDITC) solution dissolved in DMF, place in a constant temperature oscillator, and react for 10 h;

[0102] (5) Wash the reacted plastic particles 6 times repeatedly with acetone and ultrapure water;

[0103] (6) Take the newly prepared plastic particles activated by PDITC and place them in a 30mL centrifuge tube;

[0104] (7) Add 25 mL of PBS, add 4 mg of streptavidin, and place in a constant temperature shaker to react for 20 min;

[0105] (8) Centrifuge, measure the OD value of the solution at 280nm before and after the r...

Embodiment 3～6

[0107] Embodiment 3~6 prepares the kit of detection ERBB2 protein of the present invention

[0108] Monoclonal antibodies were replaced with polyclonal antibodies, genetically engineered antibodies, proteins, macromolecular organic compounds, and the rest were prepared in the same way as in Example 1 to prepare the kit for detecting ERBB2 protein of the present invention.

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Abstract

The invention discloses a kit used for detecting ERBB2 protein, and a preparation method thereof, and belongs to the field of chemiluminescence immune assay technology. The kit is capable of solving technical problems of existing ERBB2 protein ultramicro detection technology. The kit comprises: 1) an ERBB2 protein calibration material; 2) a streptavidin-coupled solid phase carrier; 3) a biotinylated ERBB2 protein ligand; 4) a 5, 10, 15- tris(4-pyridyl)-20-R-carboxylporphyrin coupling labeled ERBB2 protein ligand; and 5) a chemiluminescence substrate of 5, 10, 15-tris(4-pyridyl)-20-R-carboxylporphyrin.

Description

technical field [0001] The invention relates to the technical field of chemiluminescence immunoassay, in particular to a kit for detecting ERBB2 protein and a preparation method thereof, which combine biotin-avidin immunomagnification technology and chemiluminescence immunoassay technology. Background technique [0002] ERBB2 protein (also known as HER-2, CD340, NEU, NGL, TKR1, c-erbB-2, HER-2 / neu) is a cell membrane receptor encoded and expressed by the proto-oncogene erbB-2. Molecular weight 185kDa. In the process of scientific research and production, it is necessary to measure the change of ERBB2 protein content in the solution to monitor the experimental conditions and monitor the production process. [0003] Currently, ERBB2 protein assay methods include colloidal gold immunochromatography (GICA); Western blot (WB); enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay (CLIA); the advantage of GICA, ELISA, and WB is relatively low cost The disadvan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/532G01N33/54326G01N33/544G01N33/6803
Inventor 李勇胡庆锋常立峻徐海伟陈秀发
Owner SUZHOU HYBIOME BIOMEDICAL ENG CO LTD
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