Method for detecting 146S antigen in foot-and-mouth disease vaccine based on capillary electrophoresis method and application thereof

A foot-and-mouth disease vaccine and capillary electrophoresis technology, which is applied in the field of quantitative detection of 146S antigens of different serotypes, can solve the problems of inability to realize typing detection, low sample volume, etc., to improve market detection efficiency, good repeatability, and avoid structure damage and adsorption. effect of loss

Active Publication Date: 2020-12-04
INST OF PROCESS ENG CHINESE ACAD OF SCI
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Problems solved by technology

Although high performance liquid size exclusion chromatography has the advantages of fast detection speed, high sensitivity, low sample volume, and good repeatability, size exclusion chromatography is separated according to the size of the analyzed sample, and multivalent vaccine samples are different. Serotype 146S virus particles are similar in size, and typing detection is also impossible

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  • Method for detecting 146S antigen in foot-and-mouth disease vaccine based on capillary electrophoresis method and application thereof
  • Method for detecting 146S antigen in foot-and-mouth disease vaccine based on capillary electrophoresis method and application thereof
  • Method for detecting 146S antigen in foot-and-mouth disease vaccine based on capillary electrophoresis method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] The present embodiment provides a method for detecting the 146S antigen in the foot-and-mouth disease vaccine based on capillary electrophoresis, and the specific steps are:

[0062] (1) Preparation of injection samples:

[0063] A, prepare the aqueous phase sample of finished product foot-and-mouth disease vaccine:

[0064] Because the adjuvant is added in the finished product of the vaccine, it needs to be demulsified before the measurement; the method of demulsification is: add n-pentanol according to the volume of the vaccine stock solution, the volume ratio of the stock solution volume to n-pentanol is 9:1, mix After homogenization, let stand at 4°C for 1 hour, centrifuge at 5000 rpm for 2 minutes, collect the bottom layer solution containing the antigen as the water phase sample for injection;

[0065] B, prepare the aqueous phase sample of semi-finished product foot-and-mouth disease vaccine:

[0066] The semi-finished foot-and-mouth disease vaccine has not bee...

Embodiment 2

[0070] This example is used to demonstrate the feasibility of using capillary zone electrophoresis to detect 146S antigens of different serotypes. The method used is to analyze the charge heterogeneity of antigens of different serotypes, and the specific steps are:

[0071] The pure FMD 146S of type A, type O and Asia 1 with a concentration of 100 μg / mL was used to detect the particle size and Zeta potential of three different types of serotype virus 146S by dynamic light scattering, and three parallel tests were carried out respectively to determine The results are shown in Table 1.

[0072] Table 1

[0073] serotype Particle size (nm, n=3) Zeta potential (mV, n=3) Type A 27.3±0.2 -5.2±0.3 Type O 26.9±0.3 -10.8±0.5 Asia Type 1 27.1±0.2 -13.9±0.5

[0074] From the results in the above table, it can be seen that the particle size of different types of 146S serotypes is similar, all about 27nm, but the Zeta potentials are quite different,...

Embodiment 3

[0077] The purpose of this example is to study the influence of the background buffer salt system, detection conditions and additives on the results of capillary electrophoresis.

[0078] (1) Effect of background buffer salt type on detection results

[0079] 1. Buffer system: Select three different types of buffer systems, namely 20mM phosphate buffer, 20mM Tris-HCl buffer, and 20mM HEPES buffer, and use the resolution and characteristic peak height as parameters to investigate the effects of different buffer system types on the three serums. Effect of type 146S detection.

[0080] 2. The pH of the buffer: the pH investigation range is 7.5 to 9.0, and the specific settings are 7.5, 8.0, 8.5 and 9.0; the reason for the setting is: pH can affect the stability of 146S, and 146S is extremely unstable in a solution environment with a pH less than 7.0 And cleavage occurs; at the same time, pH will also affect the migration rate of electrophoresis, thereby affecting the separation ...

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Abstract

The invention provides a method for detecting a 146S antigen in a foot-and-mouth disease vaccine based on a capillary electrophoresis method and application thereof. The method comprises the followingsteps of introducing a water-phase sample of the foot-and-mouth disease vaccine into a capillary tube by adopting pressure sample introduction, carrying out electrophoretic separation, detecting andrecording a characteristic peak of a 146S antigen in the water-phase sample, then integrating to obtain a peak area of the characteristic peak, and then obtaining the concentration of the 146S antigenaccording to a quantitative standard curve. The method provided by the invention can be used for quantitatively detecting multiple serotype 146S antigens at the same time, can be used for detecting monovalent, bivalent or trivalent foot-and-mouth disease vaccines, has the advantages of low sample size, high sensitivity, high detection speed and the like, has important significance for realizing rapid sampling inspection of market vaccines and improving the market detection efficiency, and meanwhile, has huge application value in the aspects of product research and development and quality supervision of multivalent foot-and-mouth disease vaccines.

Description

technical field [0001] The invention belongs to the technical field of antigen quantitative detection, and in particular relates to a capillary electrophoresis method for quantitative detection of different serotype 146S antigens in monovalent, bivalent and multivalent foot-and-mouth disease oil emulsion inactivated vaccines. Background technique [0002] Foot-and-mouth disease (FMD), as a severe infectious disease, spreads quickly, spreads widely, and causes great harm, bringing huge economic losses to my country's animal husbandry industry. At present, my country adopts inactivated vaccine immunization as the main prevention and control strategy for FMD in China, and the inactivated vaccine used is mainly biphasic oil emulsion vaccine. The effective antigen that plays the most important protective role in the foot-and-mouth disease vaccine is its complete virus 146S. 146S includes seven serotypes including A, O, C, South African 1 (SAT 1), South African 2 (SAT 2), South A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
CPCG01N27/447G01N27/44704G01N27/44721G01N27/44726
Inventor 张松平杨延丽苏志国宋艳民
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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