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Kit for detecting gastric cancer antigens MG7-Ag and application thereof

A kit, mg7-ag technology, applied in the direction of measuring devices, analysis by making materials undergo chemical reactions, instruments, etc., can solve the problems of poor specificity and low detection sensitivity, and achieve high specificity, high detection sensitivity, and detection high efficiency effect

Pending Publication Date: 2020-01-14
FOURTH MILITARY MEDICAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The composition provided by the present invention includes two kinds of monoclonal antibodies that recognize different antigenic epitopes. The kit prepared by using the composition of the present invention can have both the high affinity of polyclonal antibodies and the specificity of limited epitopes of monoclonal antibodies. It overcomes the common problems of low sensitivity and poor specificity of MG7-Ag detection at present, and can effectively and conveniently carry out early diagnosis and intervention on gastric cancer, so as to reduce tumor incidence and cancer mortality

Method used

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  • Kit for detecting gastric cancer antigens MG7-Ag and application thereof
  • Kit for detecting gastric cancer antigens MG7-Ag and application thereof
  • Kit for detecting gastric cancer antigens MG7-Ag and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Preparation and use of MG7-Ag ELISA kit and MG7-Ag chemiluminescence kit method

[0064] 1.1 ELISA kit for detection of gastric cancer antigen MG7-Ag

[0065] It includes: ELISA microplates coated with MG7-Ab and MGd1-Ab; tumor marker MG7-Ag standard solution, MG7-Ag antibody horseradish peroxidase-labeled enzyme conjugate, concentrated washing solution (20X ), chromogenic solution H 2 o 2 and TMB, stop solution.

[0066] 1.2 Chemiluminescence kit for detection of gastric cancer antigen MG7-Ag

[0067] It includes: chemiluminescent microplates coated with MG7-Ab and MGd1-Ab; tumor marker MG7-Ag standard solution, MG7-Ag antibody horseradish peroxidase-labeled enzyme conjugate, concentrated washing solution (20X) , Chemiluminescent chromogen.

[0068] 1.3 Preparation method of ELISA detection kit

[0069] 1.3.1 Pre-coated 96-well microplate

[0070] The 96-well microplate is a detachable and combined 96-well transparent ELISA microplate, and the coat...

Embodiment 2

[0117] Example 2 Performance Analysis of MG7-Ag ELISA Kit

[0118] 2.1 Linear Analysis

[0119] Dilute the high-concentration sample close to the upper limit of the linear range with a low-concentration sample close to the lower limit of the linear range, mix them into test samples of at least 5 dilution concentrations (xi), test the above samples separately with the kit, and test each dilution concentration 3 times, respectively. Find the mean value (yi) of the test results for each dilution concentration. The linear regression equation was obtained with the dilution concentration (xi) as the independent variable and the mean value of the test results (yi) as the dependent variable. Computes the correlation coefficient (r) for linear regression.

[0120] The results showed that the linear range of the kit was 2ng / mL-50ng / mL, and the linear correlation coefficient r was 0.999.

[0121] 2.2 Recovery analysis (accuracy)

[0122] Calculation of the recovery rate: use the ...

Embodiment 3

[0152] Example 3 Detection of MG7-Ag ELISA Kit in Clinical Samples of Gastric Cancer, Gastritis and Normal Healthy People

[0153] The samples in this example are 9 cases of gastric cancer samples, 4 cases of atrophic gastritis and 5 cases of normal healthy human plasma samples. All samples are from Boercheng Company. Concrete method steps are as follows:

[0154] Add the MG7-Ag standard solution and the sample solution to be tested to the microwell plate respectively. 50 μL per well, and three replicate wells were made. Incubate at 37°C for 2.0 hours, pat dry with washing buffer 4 times; add antibody-enzyme conjugate, 50 μL per well, incubate at 37°C for 0.5 h, pat dry with washing buffer 4 times; 50 μL each of chromogenic substrate A solution and 50 μL chromogenic substrate B solution, develop color at 37°C in the dark for 15 minutes; termination: add 50 μL of stop solution to each well to terminate the reaction; reading: at 450 nm, use MULTIPLE FC microplate reader to r...

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Abstract

The invention provides a monoclonal antibody composition MG7-Ab, a monoclonal antibody composition MGd1-Ab and a monoclonal antibody composition CEA37-Ab which are used for detecting a gastric cancerantigen MG7-Ag. The invention further provides application of the compositions in preparation of a kit for detecting the gastric cancer antigen MG7-Ag. The kit is an ELISA kit or a chemiluminescence kit. The invention further provides application of the kit in detection of the gastric cancer antigen MG7-Ag. The kit containing two specific MG7-Ag antibodies with different antigenic epitopes is utilized for quantitatively detecting the gastric cancer antigen MG7-Ag, and the kit has the advantages of high affinity of polyclonal antibodies, specificity of limited epitopes of monoclonal antibodies,high detection efficiency, high detection sensitivity (that of the chemiluminiscence method reaches 10 pg / mL), high specificity, no need of using expensive experimental equipment, low detection costand important clinical value for diagnosis of gastric cancer.

Description

technical field [0001] The invention belongs to the field of detection of tumor markers, in particular, the invention relates to a kit for detecting gastric cancer antigen MG7-Ag and a use method thereof. Background technique [0002] Tumor protein markers are certain substances produced and released by tumor cells, which often exist in the form of metabolites such as antigens, enzymes, proteins, or peptide hormones in the patient's tumor cell tissue or in the host's body fluids and excretions. The immune profile can identify or diagnose a tumor. Tumor protein markers are mainly used clinically for the discovery of primary tumors, the screening of high-risk groups for tumors, the differential diagnosis of benign and malignant tumors, the judgment of tumor development, the observation and evaluation of tumor treatment effects, and the prediction of tumor recurrence and prognosis Wait. In recent years, new tumor markers have been continuously discovered, and inspection metho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N21/76
CPCG01N33/57446G01N21/763
Inventor 聂勇战樊代明吴开春胡思隽荆志强韩晓亮王建铭
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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