Unlock instant, AI-driven research and patent intelligence for your innovation.

Hib capsular polysaccharide purifying process

A capsular polysaccharide and process technology, applied in the field of capsular polysaccharide purification, can solve the problems of difficult removal of nucleic acid and protein, and achieve the effects of good antigenicity, qualified detection and less residual impurities

Inactive Publication Date: 2014-09-03
CHENGDU OLYMVAX BIOPHARM
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the shortcomings of the prior art, provide a safe and effective Hib capsular polysaccharide purification process that reduces harm to the human body and the environment, and solve the problem that nucleic acids and proteins are difficult to remove in the existing Hib preparation process

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
  • Hib capsular polysaccharide purifying process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Hib capsular polysaccharide purification process, it comprises the following steps:

[0024] a. Add formaldehyde solution with a final concentration of 0.8% to the fermentation broth, stir overnight at 4°C, centrifuge at 20,000rpm for 30min to remove bacteria, and collect the supernatant;

[0025] b. Filtrate the centrifuged supernatant with a 0.22um hollow fiber membrane, and collect the permeate;

[0026] c. The permeate is concentrated by ultrafiltration through the Millipore Pellicon ultrafiltration device with a 100KD membrane package, and the concentrated solution is repeatedly washed with PBS buffer until the absorbance of the concentrated solution at 280 and 260 reaches a constant value, and the concentrated washing solution is collected;

[0027] d. Add 95% ethanol to the concentrated washing solution to a final concentration of 20%, shake well and overnight at 4°C;

[0028] e. Centrifuge at 6000rpm for 30min to discard the precipitate, add 95% ethanol to the ...

Embodiment 2

[0036] Hib capsular polysaccharide purification process, it comprises the following steps:

[0037] a. Add formaldehyde solution with a final concentration of 0.8% to the fermentation broth, stir overnight at 4°C, centrifuge at 15,000rpm for 50min to remove bacteria, and collect the supernatant;

[0038] b. Filtrate the centrifuged supernatant with a 0.22um hollow fiber membrane, and collect the permeate;

[0039] c. The permeate is concentrated by ultrafiltration through the Millipore Pellicon ultrafiltration device with a 100KD membrane package, and the concentrated solution is repeatedly washed with PBS buffer until the absorbance of the concentrated solution at 280 and 260 reaches a constant value, and the concentrated washing solution is collected;

[0040] d. Add 95% ethanol to the concentrated washing solution to a final concentration of 22%, shake well and overnight at 4°C;

[0041] e. Centrifuge at 7000rpm for 20min to discard the precipitate, add 95% ethanol to the ...

Embodiment 3

[0049]Hib capsular polysaccharide purification process, it comprises the following steps:

[0050] a. Add formaldehyde solution with a final concentration of 0.8% to the fermentation broth, stir overnight at 4°C, centrifuge at 18000rpm for 60min to remove bacteria, and collect the supernatant;

[0051] b. Filtrate the centrifuged supernatant with a 0.22um hollow fiber membrane, and collect the permeate;

[0052] c. The permeate is concentrated by ultrafiltration through the Millipore Pellicon ultrafiltration device with a 100KD membrane package, and the concentrated solution is repeatedly washed with PBS buffer until the absorbance of the concentrated solution at 280 and 260 reaches a constant value, and the concentrated washing solution is collected;

[0053] d. Add 95% ethanol to the concentrated washing solution to a final concentration of 25%, shake well and overnight at 4°C;

[0054] e. Centrifuge at 8000rpm for 15min to discard the precipitate, add 95% ethanol to the su...

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

No PUM Login to View More

Abstract

The invention discloses a Hib capsular polysaccharide purifying process. The process comprises the steps that: a, a formaldehyde solution is added, the mixture is stirred overnight under a temperature of 4 DEG C; the mixture is subjected to centrifugation, and a supernatant is collected; b, the supernatant is subjected to hollow fiber filtration; c, the obtained material is subjected to ultrafiltration condensation, and a concentrated solution is washed by using PBS; d, ethanol is added to the concentrated solution, and the solution is stood overnight; e, the solution is centrifuged, and ethanol is supplemented to the supernatant; the solution is centrifuged, and a supernatant is collected; f, ethanol is added to the supernatant, and the mixture is stood; g, the mixture is centrifuged, a precipitate is collected and is dissolved by using water, the obtained solution is centrifuged, and insoluble substances are removed; h, the solution is subjected to water dialysis by using a dialysis membrane; and i, the obtained material is centrifuged and is filtered by using a filter membrane; the material is sequentially processed by using AKTAexplorer and CL-4B chromatography columns; and product of which the target peaks Kd no more than 0.5 are collected. With the process provided by the invention, difficulties that the nucleic acids and proteins are hard to in existing Hib preparation processes are solved, harms to human health and to the environment are avoided, processing difficulty is reduced, processing quality is improved, and processing stability is good.

Description

technical field [0001] The invention relates to the technical field of capsular polysaccharide purification, in particular to a Hib capsular polysaccharide purification process. Background technique [0002] Haemophilus influenzae type b (Hamephilus influenzae type b, Hib) is one of the main pathogenic bacteria that cause infectious diseases such as meningitis, sepsis and pneumonia in infants aged 3 months to 5 years old, and more than 75% of Hib infections occur In infants under 2 years of age, invasive Haemophilus influenzae type b usually causes pneumonia, cellulitis, septic arthritis, and bacteremia, with high morbidity and mortality. WHO statistics show that each year Hib can cause at least 3 million severe cases and about 386,000 deaths worldwide. Vaccination is the only public health measure to prevent the vast majority of severe cases of Hib. More than 90 countries in all regions of the world have included Hib vaccine in their routine immunization programs for chil...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00
Inventor 陈庚罗力心伍长华吴强关晓峰樊紹文
Owner CHENGDU OLYMVAX BIOPHARM