Accurate qualitative and quantitative detection method for oil adjuvant vaccine

A quantitative detection method and oil adjuvant technology, which are used in measurement devices, preparation and sampling of samples for testing, etc., can solve problems such as intractability and inability to detect water phase after demulsification, so as to improve accuracy and increase equipment maintenance. And the cost of use, the effect of improving the purity

Active Publication Date: 2017-04-26
SHANGHAI SHEN LIAN BIOMEDICAL CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Substances such as surfactants in the water phase after demulsification are recognized as the most difficult impurities in the industry, and the existing technology in the industry cannot directly detect the water phase after demulsification

Method used

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  • Accurate qualitative and quantitative detection method for oil adjuvant vaccine
  • Accurate qualitative and quantitative detection method for oil adjuvant vaccine
  • Accurate qualitative and quantitative detection method for oil adjuvant vaccine

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0048] This embodiment provides an accurate qualitative and quantitative detection method for oil adjuvant vaccines, comprising the following steps:

[0049] 1) vaccine to be tested (commercially available porcine foot-and-mouth disease vaccine, concentration is 75ug / ml) is mixed with demulsifier, organic solvent is selected acetonitrile for use in described demulsifier, acid is selected trifluoroacetic acid for use, and the volume ratio of acetonitrile and trifluoroacetic acid is 100:0.05. Take 8ml of the vaccine to be tested and put it in a centrifuge tube. Mix the vaccine and demulsifier at a volume ratio of 8:2, shake and mix well, and centrifuge at 3000G for 15 minutes at 4°C. After centrifugation, carefully extract the lower aqueous phase with a 10ml syringe.

[0050] 2) Take 0.1ml of the aqueous phase antigen sample prepared in step 1, and use HPLC to detect that the concentration of the sample is 68.8ug / ml, and the demulsification efficiency is 91.7%.

[0051] 3) Prec...

Embodiment 2

[0064] This embodiment provides an accurate qualitative and quantitative detection method for oil adjuvant vaccines, comprising the following steps:

[0065] 1) select commercially available porcine foot-and-mouth disease vaccine vaccine (vaccine concentration is 75ug / ml) and handle according to following method demulsification: vaccine to be tested is mixed with demulsifier, organic solvent is selected acetonitrile for use in described demulsifier, acid selects trifluoroacetic acid for use, The volume ratio of acetonitrile to trifluoroacetic acid is 100:0.05. Take 5ml of the vaccine to be tested and mix it with the demulsifier at a volume ratio of 5:5, shake and mix well, and centrifuge at 3000G for 15 minutes at 4°C to obtain the aqueous phase antigen sample.

[0066] 2) Take 0.1ml of the aqueous phase antigen sample prepared in step 1, and use HPLC to detect that the concentration of the sample is 67.7ug / ml, and the antigen recovery rate obtained by using this method is 90....

Embodiment 3

[0080] This embodiment provides a precise qualitative and quantitative detection method for oil adjuvant vaccines, comprising the following steps:

[0081] 1) select the commercially available porcine foot-and-mouth disease vaccine (vaccine concentration is 75ug / ml) to handle demulsification according to the following method: the vaccine to be tested is mixed with the demulsifier, and the organic solvent is selected ethanol in the described demulsifier, and the acid is selected hydrochloric acid for use, ethanol and demulsifier The volume ratio of hydrochloric acid is 100:0.33. Take 7ml of the vaccine to be tested and mix it with the demulsifier at a volume ratio of 7:3, shake and mix, and centrifuge at 3000G for 15 minutes at 4°C to obtain an aqueous phase antigen sample.

[0082] 2) Take 0.1ml of the aqueous phase antigen sample prepared in step 1, and use HPLC to detect the concentration of the sample is 70.1ug / ml, and the antigen recovery rate obtained by using this method ...

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Abstract

The invention provides an accurate qualitative and quantitative detection method for an oil adjuvant vaccine. The accurate qualitative quantitative detection method comprises the following steps of demulsifying the oil adjuvant vaccine, performing quantitative analysis on a demulsified antigen sample, meanwhile, purifying by adopting the means of gel electrophoresis, and then performing qualitative detection on the purified antigen sample; a demulsifying method comprises the following steps of mixing the oil adjuvant vaccine and a demulsifying agent and obtaining an aqueous phase antigen sample after the mixture is layered; the demulsifying agent comprises a mixture of an organic solvent and an acid, wherein the organic solvent is selected from one of acetonitrile, n-butyl alcohol, benzyl alcohol, butyl diethylene glycol and ethyl alcohol; and the acid is selected from one of hydrochloric acid, acetic acid, methanoic acid and trifluoroacetic acid. The demulsifying method provided by the invention is high in demulsifying capacity; the antigen can be enabled to be mainly distributed into the aqueous phase; the aqueous phase can be completely separated from an oil phase; the content and the quality of the effective antigen in the vaccine can be truly reflected; the demulsified antigen is separated and purified through electrophoresis; and the purity of the antigen is improved.

Description

technical field [0001] The invention relates to the technical field of foot-and-mouth disease vaccine detection, in particular to an accurate qualitative and quantitative detection method for oil adjuvant vaccines. Background technique [0002] The existing vaccine quality standards stipulate that the efficacy test must use this animal for testing. Since the country implements a 100% enhanced immunization policy, it is difficult to select susceptible animals for testing, and animal challenge has high requirements for experimental facilities (BSL3 level laboratory), time-consuming (more than one month), and large capital expenditure. If the serum neutralization test is used to select susceptible animals, it is technically difficult to exclude non-susceptible animals with cellular immunity, and the problem of irregular test data often occurs in practice, which affects the accuracy of the test. Therefore, the quality inspection of foot-and-mouth disease vaccines, especially va...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447G01N1/34G01N1/40
CPCG01N1/34G01N1/4044G01N27/447
Inventor 俞爱敏石海芳马贵军
Owner SHANGHAI SHEN LIAN BIOMEDICAL CORP
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