Method for determining content of polysaccharide of each group of meningococcus polysaccharide conjugate vaccine finished products

A meningococcal and conjugated vaccine technology, applied in the biological field, can solve the problems that it is not suitable for measuring the polysaccharide content of the conjugated vaccine finished product, the results cannot truly and accurately reflect the polysaccharide content, and the sialic acid content cannot be measured, so as to achieve enhanced detection sensitivity, High repeatability and stability, high accuracy and precision

Active Publication Date: 2012-12-05
YUXI WALVAX BIOTECH CO LTD
View PDF6 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the polysaccharide content of the stock solution of the single conjugate, the A group can be obtained by measuring the phosphorus content, and the C, Y, or W135 group can be obtained by measuring the sialic acid content, and the finished products of meningococcal polysaccharide conjugate vaccines with more than 4 groups, such as A and C , Y, and W135 groups in the finished product conjugate vaccines of groups A, C, Y, and W135 are mixed together, and groups C, Y, and W135 all contain sialic acid, so it is impossible to obtain the groups C, Y, and W135 by measuring the content of sialic acid polysaccharide content
[0003] "Methodological Validation of Quantitative Detection of Group C Meningococcal Polysaccharide Content in Quadrivalent Meningococcal Polysaccharide Vaccine by Rocket Immunoelectrophoresis" (Lin Yun et al., published in International Journal of Biological Products, Volume 31, Issue 6, December 2008 ) introduced a method for detecting the polysaccharide content of group C meningococcus in the 4-valent meningococcal polysaccharide vaccine, and solved the technical problem of determining the po

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for determining content of polysaccharide of each group of meningococcus polysaccharide conjugate vaccine finished products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Determination of Group A polysaccharide content in the finished product of Group A, C, Y, W135 meningococcal polysaccharide conjugate vaccines

[0032]The determination of the polysaccharide content of group A in the finished product of group A, C, Y, and W135 meningococcal polysaccharide conjugate vaccines was repeated three times, namely repeat 1, repeat 2, and repeat 3, and each repeat was carried out according to the following steps:

[0033] (1) Preparation of test products for inspection

[0034] ①Pretreatment: Take 12 bottles of finished meningococcal polysaccharide conjugate vaccines of groups A, C, Y, and W135 (the labeled volume is 0.5ml per bottle), and reconstitute with 6ml of water for injection (that is, reconstitute with water for injection according to the labeled volume) , take 5.0ml in an ultrafiltration cup, centrifuge ultrafiltration at 5000×g 4°C for 40min, collect the concentrate, add water for injection to 1.0ml, and obtain the ultrafil...

Embodiment 2

[0045] Example 2 Determination of the polysaccharide content of group A in the finished product of meningococcal polysaccharide conjugate vaccine of group A, C, Y, W135

[0046] Except that the operations of the following steps are different, other operations are the same as those in Embodiment 1, and will not be repeated here.

[0047] (1) Preparation of test products for inspection

[0048] ②Protease K treatment: Take 70 μl of the ultrafiltration concentrate, add 4 μl of proteinase K, 42 μl of proteinase K buffer and supplement water for injection to 420 μl, mix well and incubate at 37°C for 6 hours to obtain the test product. The amount of proteinase K added is 4 times of the protein content in the enzymolysis reaction system.

[0049] (2) Preparation of antiserum agarose gel plate

[0050] Weigh 0.1521g of agarose, add 10.0ml of pH8.6 barbiturate electrophoresis buffer, heat and swell completely, put in 56℃ water bath for 20min, add 0.3ml of meningococcal A polysacch...

Embodiment 3

[0051] Example 3 Determination of the polysaccharide content of group A in the finished product of meningococcal polysaccharide conjugate vaccine of group A, C, Y, W135

[0052] Except that the operations of the following steps are different, other operations are the same as those in Embodiment 1, and will not be repeated here.

[0053] (1) Preparation of test products for inspection

[0054] ②Protease K treatment: Take 70 μl of the ultrafiltration concentrate, add 8 μl of proteinase K, 42 μl of proteinase K buffer and make up to 420 μl with water for injection, mix well and incubate at 37°C for 8 hours to obtain the test product. The amount of proteinase K added is 8 times of the protein content in the enzymolysis reaction system.

[0055] (2) Preparation of antiserum agarose gel plate

[0056] Weigh 0.1522g of agarose, add 10.0ml of pH8.6 barbiturate electrophoresis buffer, heat and swell completely, put in 56℃ water bath for 30min, add 0.3ml of meningococcal A polysac...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Protein concentrationaaaaaaaaaa
Login to view more

Abstract

The invention relates to a method for determining the content of the polysaccharide of each group of meningococcus polysaccharide conjugate vaccine finished products and belongs to the technical field of biology. The method comprises the following steps of: treating by using protease K during preparation of a detection product sample, wherein the total size of an enzymolysis reaction system is 420 mu l; and by using the total size as reference, adding the protease K, a protease K buffer solution which occupies one tenth of the total size and an ultra-filtration concentrated solution which occupies one sixth of the total size, uniformly mixing the protease K, the protease K buffer solution and the ultra-filtration concentrated solution, and then incubating, wherein the addition amount of the protease K is two to eight times of the content of proteins in the enzymolysis reaction system. By adoption of the method, influence of each group of conjugate carrier proteins on immunoelectrophoresis is eliminated, an appropriate detection system for detecting the polysaccharide of each of more than four groups of meningococcus polysaccharide conjugate vaccine finished products is established, and the content of the polysaccharide of each of more than four groups of meningococcus polysaccharide conjugate vaccine finished products can be accurately and quickly determined. The method has the characteristics of high durability, high accuracy and high precision. A method for evaluating the quality of meningococcus polysaccharide conjugate vaccine is established.

Description

technical field [0001] The invention belongs to the field of biotechnology, and more specifically relates to a method for determining the content of polysaccharides in each group of finished meningococcal polysaccharide conjugate vaccines. Background technique [0002] The polysaccharide (antigen) content of the finished product of meningococcal polysaccharide conjugate vaccine is one of the basic conditions to ensure the effectiveness of the vaccine. For the polysaccharide content of the stock solution of the single conjugate, the A group can be obtained by measuring the phosphorus content, and the C, Y, or W135 group can be obtained by measuring the sialic acid content, and the finished products of meningococcal polysaccharide conjugate vaccines with more than 4 groups, such as A and C , Y, and W135 groups in the finished product conjugate vaccines of groups A, C, Y, and W135 are mixed together, and groups C, Y, and W135 all contain sialic acid, so it is impossible to obta...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N33/68G01N33/561
Inventor 黄镇吴凯樊会兰张胜祥马波周燕美李如彪包南艳杨志伟周进宝刘国秀钱红兵
Owner YUXI WALVAX BIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap