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Fast qualitative and quantitative detection method of oil adjuvant vaccine

A quantitative detection method and quantitative detection technology, applied in the field of foot-and-mouth disease synthetic peptide vaccine detection, can solve the problems of not responding well to the effective antigen content, and there is no universal and accurate method for quantitative detection, so as to achieve short time consumption, improve detection sensitivity, The effect of easy operation

Active Publication Date: 2017-05-31
SHANGHAI SHEN LIAN BIOMEDICAL CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Can not reflect the effective antigen content in synthetic peptide vaccine against foot-and-mouth disease
[0006] At present, there is no universal and accurate method for the quantitative detection of antigen content in synthetic peptide vaccines for FMD

Method used

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  • Fast qualitative and quantitative detection method of oil adjuvant vaccine
  • Fast qualitative and quantitative detection method of oil adjuvant vaccine
  • Fast qualitative and quantitative detection method of oil adjuvant vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] This embodiment provides a rapid qualitative and quantitative detection method for oil adjuvant vaccines, specifically using the following steps:

[0045] 1) The commercially available porcine foot-and-mouth disease synthetic peptide vaccine (vaccine antigen concentration is 50 μ g / mL) is demulsified according to the following method:

[0046] Take 10ml of the vaccine to be tested and mix it with n-butanol at a volume ratio of 1:1, add 50mg of histidine, shake and mix well, and centrifuge at 3000r / min for 15 minutes at 4°C, and carefully extract the lower layer of water with a 10ml syringe after centrifugation Phase, that is, the aqueous phase antigen sample.

[0047] The commercially available pig foot-and-mouth disease synthetic peptide vaccine used in this embodiment is compared with the theoretical antigen concentration standard, and the peak position of its HPLC detection chromatogram sample is integrated, and its integral information is shown in Table 1. In the f...

Embodiment 2

[0064] This embodiment provides a rapid qualitative and quantitative detection method for oil adjuvant vaccines, specifically using the following steps:

[0065] 1) The commercially available porcine foot-and-mouth disease synthetic peptide vaccine (vaccine antigen concentration is 50 μ g / mL) is demulsified according to the following method:

[0066] Take 10ml of the vaccine to be tested and mix it with n-butanol at a volume ratio of 1:1, add 10mg of phenylalanine, shake and mix well, centrifuge at 3000r / min for 15 minutes at 4°C, and carefully extract the lower layer with a 10ml syringe after centrifugation Aqueous phase, namely the aqueous phase antigen sample.

[0067] The foot-and-mouth disease vaccine was demulsified by the method of this example, and the demulsification efficiency was 88.4%.

[0068] 2) Competitive ELISA detection

[0069] 2.1 Antigen coating: 3 μg / mL artificially synthesized FMD antigen, 100 μL per well, was immobilized onto the ELISA plate;

[0070]...

Embodiment 3

[0082] This embodiment provides a rapid qualitative and quantitative detection method for oil adjuvant vaccines, specifically using the following steps:

[0083] 1) The commercially available porcine foot-and-mouth disease synthetic peptide vaccine (vaccine antigen concentration is 50 μ g / mL) described in embodiment 1 is demulsified according to the following method:

[0084] Take 10ml of the vaccine to be tested and mix it with n-butanol at a volume ratio of 1:1, add 200mg of proline to each tube, vortex and mix well, centrifuge at 3000r / min for 15 minutes at 4°C, and carefully extract with a 10ml syringe after centrifugation The lower aqueous phase is the aqueous phase antigen sample.

[0085] The foot-and-mouth disease vaccine was demulsified by the method of this example, and the demulsification efficiency was 95.6%.

[0086] 2) Competitive ELISA detection

[0087] The competition ELISA method used was the same as in Example 2. The actual content of the antigen in the s...

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Abstract

The invention provides a fast qualitative and quantitative detection method of an oil adjuvant vaccine. The method comprises the following steps: carrying out demulsification on a synthetic peptide vaccine for foot-and-mouth disease, and carrying out qualitative and quantitative detection on an antigen sample, subjected to the demulsification, by using a competitive enzyme-linked immunosorbent assay (ELISA), wherein a demulsification method comprises the steps of mixing the oil adjuvant vaccine with n-butyl alcohol, then adding a competitive agent into the mixture, evenly mixing, and centrifuging to obtain the antigen sample. According to the method, an antigen in the oil adjuvant vaccine is released into a water phase by adding the competitive agent and a surface active agent to compete for an antigen binding site, so that the recovery rate of the antigen in the water phase is greatly increased, the sample can be measured without being purified, and the detection time is shortened. According to the method, the antigen to be detected is firstly enabled to react with an antibody having a known concentration, so that the antibody is firstly and completely neutralized with the antigen; after that, a neutralized solution is enabled to react with the antigen which is absorbed on an ELISA plate in an immobilized way, so that the concentration of the antigen to be detected is measured, and a standard curve slope obtained by using the detection method provided by the invention is greater than that obtained by using the conventional indirect competition method; therefore, the detection sensitivity is greatly improved.

Description

technical field [0001] The invention relates to the technical field of detection of foot-and-mouth disease synthetic peptide vaccines, in particular to a rapid 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 vacci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983
Inventor 刘自立马贵军石海芳俞爱敏姬明放
Owner SHANGHAI SHEN LIAN BIOMEDICAL CORP
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