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

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...

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 polysaccharide content of each group of meningococcal polysaccharide vaccines of groups A, C, Y, and W135. However, this method is not suitable for the determination of the polysaccharide content of each group in the finished conjugated vaccine, because experiments have proved that the carrier protein of each group of conjugates in the conjugated vaccine will affect the rocket immunoelectrophoresis, resulting in results that cannot truly and accurately reflect the polysaccharide content, so the determination The polysaccharide conjugate vaccine products of more than 3 groups include A, C, Y, and W135 meningococcal polysaccharide conjugate vaccines. The polysaccharide content of each group of the finished polysaccharide conjugate vaccine is a technical difficulty. At present, there is no relevant method for the determination of the polysaccharide content of each group of the polysaccharide conjugate vaccine product. According to the report, it is necessary to establish a method to accurately determine the polysaccharide content of each group A, C, Y, W135, etc. in the finished polysaccharide conjugate vaccine.

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