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Quantitative detection method of rabies virus inactivated antigen

A quantitative detection method and a technology for inactivated antigens, which are applied in the field of quantitative detection of rabies virus inactivated antigens, and can solve the problems of difficulty in rapid quantification of rabies virus.

Active Publication Date: 2021-09-14
CHINA ANIMAL HUSBANDRY IND
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Problems solved by technology

According to the structural characteristics of rabies virus, the present invention designs a high-performance liquid chromatography detection method using gel column separation, combines sample processing, preparation of standard products and optimization of detection conditions, establishes an efficient quantitative method for rabies inactivated antigen, and solves the problem Key technical issues that are difficult to quickly quantify rabies virus in actual production

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  • Quantitative detection method of rabies virus inactivated antigen
  • Quantitative detection method of rabies virus inactivated antigen
  • Quantitative detection method of rabies virus inactivated antigen

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Experimental program
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Effect test

Embodiment 1

[0049] 1. Instruments and reagents

[0050] Shimadzu high performance liquid chromatograph LC20A, equipped with UV detector.

[0051] Preparation of mobile phase: Take disodium hydrogen phosphate dodecahydrate, sodium dihydrogen phosphate dihydrate and anhydrous sodium sulfate, dissolve in ultrapure water, set the volume to 1000mL, and prepare a solution containing 10mM disodium hydrogen phosphate, 40mM phosphoric acid Sodium dihydrogen and 0.1M sodium sulfate, pH 7.2 buffer, 0.45μm membrane filter. The gel chromatography column is Thermo Bio Basic SEC-1000 or TSKgel G4000SWXL or TSKgel G6000 PWXL; the flow rate is 0.6ml / min.

[0052] Sample to be tested: rabies virus suspension culture inactivated antigen

[0053] Loading volume: 50μl

[0054] 2. Rabies virus standard

[0055] Purchase the national standard product of rabies vaccine (article number: 250009) from the Central Inspection Institute.

[0056] figure 1 It is the HPLC detection result of the rabies virus stand...

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Abstract

The invention discloses a quantitative detection method of rabies virus inactivated antigen. The method comprises: at first using the SDS-PAGE method and the TEM method to qualitatively identify the target peak; then using a protein quantification kit to measure the concentration of the standard substance; serially diluting the standard substance, and detecting it on a chromatograph with a gel chromatographic column According to the absorption peak of the standard substance at 280nm, the linear regression equation of concentration and peak area is established; the concentration of rabies virus inactivated antigen in the test sample is calculated according to the linear regression equation and the peak area of ​​the test sample. The method of the invention can be used for the quality control of rabies virus production, has the advantages of rapidity, stability and good repeatability, can guide the production of vaccines, and plays an important role in improving the quality of vaccines.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a quantitative detection method for viruses, in particular to a quantitative detection method for rabies virus inactivated antigens. Background technique [0002] Rabies virus is a zoonotic, acute and highly lethal infectious disease caused by Rhabdoviridae genus Lyssavirus. According to the analysis and identification of rabies virus N protein monoclonal antibody and G protein series, rabies viruses around the world are mainly divided into 4 serotypes. Among them, serotype 1 is the well-known rabies virus with the widest geographical distribution and the greatest threat to the health of people in the world, including wild viruses and fixed laboratory strains (such as SAD). With the research progress of modern molecular biology, referring to the similarity rate of the N-terminal 500 nucleotides of the rabies virus nucleoprotein (N protein) gene, the lyssavirus is divided into 7 genotyp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/8831
Inventor 宋芳肖进张蕾董春娜齐鹏栗利芳刘新月巴利民侯旭李玲李静张文亮李鹏宇
Owner CHINA ANIMAL HUSBANDRY IND
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