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Method for measuring antigen protein in serum by chemiluminescence protein chips and kit

A chemiluminescence and protein chip technology, applied in the field of serum antigen detection, can solve the problems of long detection time, high cost, large blood volume, etc., and achieve the effect of reducing detection cost, low cost and high sensitivity

Inactive Publication Date: 2014-05-28
BEIJING YOUAN HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above detection methods have the problems of large blood volume, long detection time, and high cost.

Method used

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  • Method for measuring antigen protein in serum by chemiluminescence protein chips and kit
  • Method for measuring antigen protein in serum by chemiluminescence protein chips and kit
  • Method for measuring antigen protein in serum by chemiluminescence protein chips and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1. Protein chip preparation and use process

[0057] Reagents used in the experiment: mouse monoclonal antibody AFP (Shenzhen Faipeng Company); aldehyde-based chip (Shanghai Biopro Company); biotin-labeled rabbit-derived primary antibody (Abcam Company, USA); HRP-labeled streptavidin ( American abcam company); HRP substrate luminescent fluid (US millipore company), chemical imager (developed by the Academy of Military Medical Sciences).

[0058] In a preferred embodiment of the present invention, the mouse monoclonal antibody AFP (Shenzhen Faipeng Company) was spotted on the chip in turn, and the sample was spotted four times at a spotting concentration of 2 mg / ml to form a detection spot.

[0059] 10% bovine serum albumin (BSA) was used as a negative control, and the sample was also spotted four times to form a control spot. The chip is an aldehyde-based chip (Shanghai BioPro Company), each chip contains 10 detection squares, each square detects one serum, and...

Embodiment 2

[0067] Embodiment 2. Establishment of the detection method of the present invention

[0068] Step 1: Develop a Standard Curve

[0069] Use the purchased AFP antigen (abcam company in the United States), set to different concentration gradients, 80ng / ml, 40ng / ml, 20ng / ml, 10ng / ml, 5ng / ml, 2.5ng / ml, 1.25ng / ml, 0.625ng / ml.

[0070] Adopt the operation flow in the embodiment 1 and protein chip to detect each concentration gradient of AFP standard substance, scanning result

[0071] Such as image 3 .

[0072] Table 1. Standard curve detection data results

[0073]

[0074]

[0075] The test results are drawn as a standard curve, with the concentration of the standard as the abscissa and the pixel value as the ordinate, and the standard curve is drawn on the coordinate paper; the corresponding concentration is found from the standard curve according to the pixel value of the sample; then multiplied by the dilution Multiple; or use the concentration and OD value of the s...

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Abstract

The invention relates to a method for measuring antigen protein in serum by chemiluminescence protein chips and a kit, which belongs to the medical detection technology. The method is characterized in that a chemiluminescence technology, a standard curve and a protein chip technology are combined for performing quantitative determination of certain antigen protein substance in serum, the high specialty and sensitivity of the chemiluminescence technology and the standard curve are employed for realizing the quantitative determination; the protein chip technology can realize simultaneous detection and analysis of several samples, so that the detection time consumption of a whole set ofe sample is greatly reduced, and the method has the characteristics of reliability, accuracy and low cost; and the method is suitable for general analysis of common antigen protein level in serum and quantitative analysis of great disease antigen marks, and the method is an medical auxiliary detection method having great meaning. The invention also provides a matched kit based on the method.

Description

technical field [0001] The invention relates to a serum antigen detection technology, in particular to a chemiluminescence protein chip method and a kit for antigen-like proteins in serum. Background technique [0002] Hepatocellular carcinoma is one of the most common malignant tumors in the world. my country is a big country with hepatitis B, and the incidence of liver cancer accounts for 55% of the global total. Nearly 80% of the patients are in the middle and late stage when they are diagnosed. The postoperative recurrence and metastasis rate is high, and the long-term survival rate over 5 years is only about 10%. Early diagnosis of liver cancer is the best and effective way to improve the long-term survival rate of patients. [0003] In the field of early diagnosis of liver cancer, serum tumor molecular markers have been widely concerned. At present, basic research has found that more than 100 markers are related to liver cancer. AFP is still the best marker for the q...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N21/64G01N21/76
CPCG01N33/542G01N33/57438G01N33/6893
Inventor 李宁张爱英张永宏王升启
Owner BEIJING YOUAN HOSPITAL CAPITAL MEDICAL UNIV
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