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Agglutinant competitive process liver cancer a-fetoglobulin heteroplasmon AFP-L3 quantitative test reagent kit

A technology for AFP-L3 and alpha-fetoprotein, which is applied in the field of kits for detecting the content of alpha-fetoprotein heterogenous AFP-L3, can solve the problems of difficulty in distinguishing benign and malignant lesions, high technical requirements, complicated operation and the like, and achieves the method Simple application, high sensitivity, overcoming the effect of cumbersome operation steps

Inactive Publication Date: 2006-05-17
BEIJING WEILONG ARRAY BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] 2) Differentiate between liver cancer and benign liver disease AFP is often elevated in patients with primary liver cancer, but many benign liver diseases can also have elevated AFP, sometimes it is difficult to distinguish benign and malignant lesions based on AFP results alone
[0012] However, for a long time, only a few clinical laboratories in China can measure AFP heterogeneity, and only a few cities can meet the requirements for clinical determination of AFP heterogeneity
So far, the heterogeneous body determination method (currently the main method is electrophoresis separation affinity adsorption method), has high technical requirements, cumbersome operation, and expensive reagents

Method used

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  • Agglutinant competitive process liver cancer a-fetoglobulin heteroplasmon AFP-L3 quantitative test reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Production of AFP-L3 ELISA Quantitative Detection Kit

[0053] A kind of AFP-L3 ELISA quantitative detection kit (96 people), its composition comprises:

[0054] One bottle of AFP-L3 negative and one positive control;

[0055] AFP monoclonal antibody coated plate (96 wells) 1 piece;

[0056] Horseradish peroxidase (HRP)-labeled AFP-L3 monoclonal antibody 1 bottle, 6ml / bottle;

[0057] Lentil agglutinin (LCA) solution 1 bottle

[0058] 1 bottle of blank buffer

[0059] 1 bottle of AFP standard

[0060] 1 bottle each of substrate solution A and chromogenic solution B, each 5ml / bottle;

[0061] 1 bottle of reaction termination solution, 5ml / bottle;

[0062] Wash buffer (20X concentrated) 1 bottle, 30ml / bottle.

[0063] The specific operation is as follows:

[0064] 1. Prepare AFP-negative and positive controls:

[0065] 1) Serum collection: Obtain the serum of healthy normal people and ascites of liver cancer patients from the hospital or blood bank, an...

Embodiment 2

[0109] Example 2: Use of AFP-L3 Quantitative ELISA Kit

[0110] 1. Washing buffer preparation: dilute the concentrated washing buffer provided in the kit with distilled water 20 times.

[0111] 2. Antigen-antibody reaction: Add 50 μl of positive or negative quality control serum, or serum samples to be tested, to the microwells of the antibody-coated plate provided in the kit, and incubate in a water bath at 37°C for 30 minutes, wash the plate 5 times, Add lectin solution and control solution to each double well, incubate in 37°C water bath for 30 minutes, shake off the liquid, add HRP-monoclonal antibody solution to each well, 50ul per well, incubate in 37°C water bath for 30 minutes. Repeat the plate washing operation 5 times, 3 minutes each time.

[0112] 3. Color reaction: Add 50 μl each of substrate solution A and TMB chromogenic solution B to each well in turn, incubate in a 37°C water bath for 10 minutes, and then add 50ul of reaction termination solution to each well ...

Embodiment 3

[0121] Example 3: Detection of positive samples of liver cancer patients by the AFP-L3 ELISA quantitative detection kit of the present invention

[0122] In order to judge the coincidence rate between the AFP-L3 ELISA quantitative detection kit of the present invention and the clinical liver cancer detection results, the present invention is used for detection. Clinical research data of Ditan Hospital: 100 known positive specimens of primary liver cancer, 50 healthy serum samples from physical examination, and 100 liver cirrhosis serum samples were tested with AFP-L3 enzyme-linked immunosorbent assay kit. The results are shown in Table 1.

[0123] serum source

[0124] The results of this experiment show that the coincidence rate of the present invention for primary liver cancer is as high as 82%, and the specificity for liver cirrhosis is as high as 95%.

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Abstract

The present invention provides a monoclonal antibody, It is obtained by utilizing screened hybridoma with monoclonal antibody which can be combined with AFP heteroplasmon glycochain and can be competed with dolichos agglutinin site and cloning it. Said invention also provides kit containing said monoclonal antibody, its preparation method and its application. Said kit can be used as index for early diagnosis of liver cancer.

Description

technical field [0001] The invention belongs to the field of biotechnology, and more specifically, the invention provides an alpha-fetoprotein monoclonal antibody whose binding site is near the sugar chain of the alpha-fetoprotein heterogeneity, which can be masked by the competitive binding of lentil lectin , and provide a kit for detecting heterogeneous AFP-L3 content of alpha-fetoprotein developed by using the monoclonal antibody, its preparation method and application. The kit can be used to quickly and accurately diagnose the occurrence of liver cancer at an early stage. Background technique [0002] Worldwide, hepatocellular carcinoma is the fourth most common cancer and the third most common cause of cancer-related death. Risk factors include chronic hepatitis and cirrhosis of the liver caused by the hepatitis B virus and hepatitis C virus. The clinical efficiency of screening and screening depends on early diagnosis for effective treatment. [0003] AFP-L3 is the ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531
Inventor 林长青
Owner BEIJING WEILONG ARRAY BIOLOGICAL TECH
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