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Method of detecting molecular weight of pneumococcal capsular poly-saccharides

A technology of capsular polysaccharide and pneumococcus, applied in the biological field, can solve the problems of poor repeatability, time-consuming, labor-consuming, etc., and achieve the effects of high accuracy, good repeatability and improved accuracy

Active Publication Date: 2020-01-31
BEIJING ZHIFEI LVZHU BIOPHARM +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] As of January 2019, a 23-valent pneumococcal polysaccharide vaccine has been launched in China, and many vaccine manufacturers are developing pneumococcal polysaccharide vaccines and pneumococcal polysaccharide conjugate vaccines, but the third part of the Chinese Pharmacopoeia (2015 edition) has not yet included 23-valent pneumococcal polysaccharide vaccines. The manufacturing and verification regulations of pneumococcal polysaccharide vaccines, the vaccine inspection regulations included in the European Pharmacopoeia version 7.0 stipulate that the partition coefficients of various types of PnPs samples in chromatographic columns should be determined by gel size-exclusion chromatography (SEC) (K D ), and stipulated the K of each monovalent polysaccharide D However, this method has disadvantages such as time-consuming, labor-intensive, and poor repeatability.

Method used

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  • Method of detecting molecular weight of pneumococcal capsular poly-saccharides
  • Method of detecting molecular weight of pneumococcal capsular poly-saccharides
  • Method of detecting molecular weight of pneumococcal capsular poly-saccharides

Examples

Experimental program
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Effect test

Embodiment 1

[0046] 1) Instruments and reagents

[0047]JA2003 electronic balance was purchased from Shanghai Sunny Hengping Scientific Instrument Co., Ltd.; DV2T viscometer was purchased from BROOK FIELD company, rotor model: CPA-40Z; e2695 high-efficiency liquid phase system pump, Waters 2414 differential refraction detector were purchased from Waters company; The high-performance liquid chromatography column is TSK PWXL Guard protection column and TSK G5000PWXL (7.8mmID×30cm) analysis column purchased from Japan TOSOH Corporation.

[0048] Reagents: Sodium dihydrogen phosphate dodecahydrate, disodium hydrogen phosphate dihydrate, sodium chloride, and dextran standards were purchased from Waters and American Polymer Standards Corporation.

[0049] Preparation of 0.1mol / L phosphate buffer solution: Weigh 0.4g sodium dihydrogen phosphate dodecahydrate, 6.0g disodium hydrogen phosphate dihydrate, 9.0g sodium chloride, add deionized water to 1000g.

[0050] 2) Preparation of samples for ser...

Embodiment 2

[0057] 1) Instruments and reagents

[0058] JA2003 electronic balance was purchased from Shanghai Sunny Hengping Scientific Instrument Co., Ltd.; DV2T viscometer was purchased from BROOK FIELD company, rotor model: CPA-40Z; e2695 high-efficiency liquid phase system pump, Waters 2414 differential refraction detector were purchased from Waters company; The high-performance liquid chromatography column is TSK PWXL Guard protection column and TSK G5000PWXL (7.8mmID×30cm) analysis column purchased from Japan TOSOH Corporation.

[0059] Reagents: Sodium dihydrogen phosphate dodecahydrate, disodium hydrogen phosphate dihydrate, sodium chloride, and dextran standards were purchased from Waters and American Polymer Standards Corporation.

[0060] Preparation of 0.1mol / L phosphate buffer solution: Weigh 0.4g sodium dihydrogen phosphate dodecahydrate, 6.0g disodium hydrogen phosphate dihydrate, 9.0g sodium chloride, add deionized water to 1000g.

[0061] 2) Preparation of 33F serotype P...

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Abstract

The invention discloses a method of detecting molecular weight of pneumococcal capsular poly-saccharides (PnPs) by an HPSEC-RI method. The method comprises the steps of preparing a mobile phase, precisely weighing the PnPs, adding the mobile phase, performing dissolution, performing dilution by the mobile phase, detecting the viscosity of the PnPs to allow a viscosity value of a diluted sample tobe within a certain range, performing filtering as a test sample solution, selecting a plurality of glucose standard substances with different molecular weight, respectively performing dilution by themobile phase, performing sample injection, performing analysis by liquid chromatography, establishing a molecular weight standard curve and a regression equation, detecting and recording a test sample under the same test condition, and calculating the molecular weight of the sample by applying the established standard curve and the established regression equation. A buffer system and a sample processing method are high in precision and good in repeatability.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for detecting the molecular weight of pneumococcal capsular polysaccharide. Background technique [0002] Currently, there are 23-valent pneumococcal polysaccharide vaccines and 7-valent / 13-valent pneumococcal polysaccharide conjugate vaccines for the prevention of Streptococcus pneumoniae infection. Pneumococcal capsular polysaccharides (Pneumococcal capsular poly-saccharides, PnPs) is the main active ingredient of the vaccine, studies have shown that the molecular weight of PnPs directly affects the immunogenicity of the polysaccharide vaccine; Quality is also an important indicator of quality control. [0003] As of January 2019, a 23-valent pneumococcal polysaccharide vaccine has been launched in China, and many vaccine manufacturers are developing pneumococcal polysaccharide vaccines and pneumococcal polysaccharide conjugate vaccines, but the third part of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/86
CPCG01N30/02G01N30/06G01N30/8631
Inventor 马可朱卫华宗向坤张霁赵林飞胡月凤杜琳
Owner BEIJING ZHIFEI LVZHU BIOPHARM
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