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Test method for hog cholera virus E2 protein quantification

A swine fever virus and detection method technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve the effect of low requirements for personnel and equipment

Inactive Publication Date: 2016-01-13
TECON BIOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no institutions and scholars in China that use monoclonal antibodies to capture the E2 protein of classical swine fever virus and combine it with the BCA method for quantitative and qualitative analysis. This method has simple steps, saves time and is easy to operate

Method used

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  • Test method for hog cholera virus E2 protein quantification
  • Test method for hog cholera virus E2 protein quantification
  • Test method for hog cholera virus E2 protein quantification

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Embodiment 1

[0012] Embodiment 1, the quantitative detection method of classical swine fever virus E2 protein, one, preparation and purification for the monoclonal antibody of classical swine fever virus E2 protein: 1. the hybridoma cell that will prepare monoclonal antibody is cultivated according to conventional culture mode, after culturing Healthy cells were centrifuged and adjusted to 1.0 x 10 6 / ml; ②The density-adjusted hybridoma cells were injected into the peritoneal cavity of Bacb / c mice, and the ascites in the peritoneal cavity of the mice was extracted 14 days later; ③After preliminary purification with saturated ammonium sulfate, further purification was performed with MelonGelIgGSpin Purification Kit; ④BCA reagent was used The box quantifies the purified monoclonal antibody, and it is ready for use after quantification.

[0013] 2. Optimization of coating, coating concentration, and concentration of CSFV E2 protein monoclonal antibody to be tested: ①Use the coating buffer to ...

Embodiment 2

[0015] Embodiment 2, the verification of specificity: get and express classical swine fever virus E2 protein, porcine circovirus (II) on insect cells, express porcine circovirus Cap protein on insect cells, porcine pseudorabies virus, porcine parvovirus, bovine The cell culture of viral diarrhea virus was tested, and the results showed that except for the positive expression of classical swine fever virus E2 protein on insect cells, the others were all negative, which proved that the coated monoclonal antibody had good specificity.

[0016] Quantitative accuracy verification of CSFV E2 protein: Randomly select 5 samples of CSFV E2 recombinant baculovirus expressed on insect cells for detection of CSFV E2 protein after dilution of stock solution, 1 / 2, and 1 / 4, respectively. The coated monoclonal antibody was detected by high-performance liquid chromatography, and the 1 / 2 and 1 / 4 diluted test results of the same sample were restored to the stock solution for statistical analysis....

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Abstract

The invention belongs to the technical field of improvement, research and development of biological test methods and particularly relates to a test method for hog cholera virus E2 protein quantification. The test method comprises the first step of preparing and purifying a hog cholera virus E2 protein monoclonal antibody, the second step of optimizing peridium and peridium concentration of the hog cholera virus E2 protein monoclonal antibody and the concentration of hog cholera virus E2 protein to be tested and the third step of determining the critical value and the test result. Compared with a conventional method, the qualitative process for quantification through the method requires about 2 hours; on the premise of not influencing the quantification result, the requirements for personnel, instruments and equipment are not high, time and labor are saved, and a novel method is provided for hog cholera virus E2 protein quantification.

Description

technical field [0001] The invention belongs to the technical field of improvement and research and development of biological detection methods, in particular to a quantitative detection method for classical swine fever virus E2 protein. Background technique [0002] Currently, methods for quantifying CSFV E2 protein include gel imaging software analysis and high performance liquid chromatography. The quantitative and qualitative analysis of CSFV E2 protein by polyacrylamide gel electrophoresis (SDS-PAGE) and Western-blot by gel imaging software analysis method takes a long time; high-performance liquid chromatography can quickly analyze CSFV Quantification of E2 protein, but the requirements for equipment and operators are high, and immunological analysis (qualitative) cannot be performed; while the quantitative and qualitative analysis of E2 protein of classical swine fever virus using monoclonal antibody is based on the principle of BCA protein quantitative method , The ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577G01N30/02
CPCG01N33/56983G01N30/02G01N33/577
Inventor 李俊辉赵毅豆智华王遵宝张先锋李延涛贺笋郑侃
Owner TECON BIOLOGY CO LTD
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