Polyvalent bacteria capsule polysaccharide-protein conjugate combined vaccine

A technology of capsular polysaccharides and combined vaccines, applied in the field of combined vaccines, to achieve the effect of reducing the number of vaccinations, relieving pain, and enhancing immunogenicity

Active Publication Date: 2005-12-21
BEIJING ZHIFEI LVZHU BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, there are exotoxin A, P6 and some high-molecular outer membrane proteins of Pseudomonas aeruginosa, as well as unrelated proteins

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Extraction of capsular polysaccharide of meningococcus group A, C, Y, W135 and capsular polysaccharide of Haemophilus influenzae type b

[0029] Proper medium can be selected for the production of polysaccharide vaccines. The liquid medium used for production does not contain components that can form precipitates with cetyltrimethylammonium bromide. The culture medium does not contain harmful or other allergen substances.

[0030] After inoculating the strains in a suitable medium, harvest after the logarithmic production phase or early in the stationary phase. It is advisable to add formaldehyde solution to the culture to sterilize or heat to sterilize, so as to ensure the safety of sterilization and not damage the polysaccharide of the bacteria. Centrifuge the sterilized single harvest (or combined harvest) to remove bacteria, collect the supernatant, concentrate the harvested supernatant to 1 / 8 of the original volume with an ultrafiltration membrane with a molecula...

Embodiment 2

[0035] Lysis and purification of capsular polysaccharide

[0036] Dissolve 5 g of group A (or group C, Y, W135) Neisserial meningitidis capsular polysaccharide (or capsular polysaccharide of Haemophilus influenzae type b, Hib) in 10 liters of 1 / 10 saturated sodium acetate buffer solution (pH5.5-6.0, adjust pH with acetic acid, room temperature), stir at room temperature, fully dissolve for 8-12 hours, until the solution is clear and transparent, add 50mM sodium acetate buffer (pH 5.5- 6.0) 38.33 liters, heat the circulating water in the jacket of the stainless steel reaction tank, maintain the temperature of the polysaccharide solution in the tank at 55-56 ° C, stir for 0.5 hours, add 1670 ml of 30% hydrogen peroxide to the tank, and crack the polysaccharide. The stirring speed is 100 rpm.

[0037] The polysaccharide cracking process is monitored by tracking the molecular weight of the polysaccharide. The molecular weight is determined by Shimadzu 20AT HPLC TSK3000W liquid ch...

Embodiment 3

[0040] Coupling and Purification of Polysaccharide and Tetanus Toxoid

[0041] 1. Activate polysaccharide

[0042] Take 400mg (4mg / ml, 100ml) of meningococcal capsular polysaccharide (or Hib) polysaccharide of group A (or group C, group Y, group W135), adjust the pH to 10.8 with 0.5M NaOH, add 200mg of cyanogen bromide, and use 0.5M NaOH to maintain the pH at about 10.8±0.5 for 1 hour (22°C). Then use 0.5M HCl to adjust the pH to 8.8, add 1450mg adipic hydrazide (ADH), and react for 6 minutes; use 0.5M NaOH to adjust the pH to 8.5, and maintain the pH within the range of 8.5±0.5 for 15 minutes; Stir gently for 12 hours. Dialyze with pre-cooled 0.05M NaCl solution for 48 hours (4-8°C), during which time the solution was changed 5 times (or ultrafiltration with 3KD polyethersulfone ultrafiltration membrane). Filter the activated polysaccharide-AH derivative with a 0.45 μm filter membrane.

[0043] 2. Coupling of activated polysaccharides and tetanus toxoid (TT) (operated und...

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PUM

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Abstract

The present invention relates to a polyvalent bacterial capsule polysaccharide-protein conjugate combined vaccine preparation, in particular, it is a combined vaccine containing group A, group C, group Y and group W135 epidemic cerebrospinal meningitis coccal capsule polysaccharide-protein conjugate and b type haemophilus influenzal capsule polysaccharide-protein conjugate.

Description

Technical field: [0001] The present invention relates to a combined vaccine preparation of polyvalent bacterial capsular polysaccharide and protein covalent conjugate, especially a kind of capsular polysaccharide-protein containing epidemic cerebrospinal meningococcus of group A, group C, group Y and group W135 Conjugate and Haemophilus influenzae type b capsular polysaccharide-protein conjugate vaccine. Background technique: [0002] Meningococcal meningitis is a human infectious disease with a long history. It is endemic in a very wide area and spreads all over the world. It has not been effectively controlled so far. The causative agent of the disease, Neisseria meningitidis, only infects humans, and is directly transmitted from person to person through droplets or secretions. Usually, this contact makes the other person a healthy carrier, depending on age and environment. Different, 10-50% of people can become carriers. When the human body's resistance is weakened and ...

Claims

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Application Information

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IPC IPC(8): A61K39/02
Inventor 孔健蒋先敏肖丽君
Owner BEIJING ZHIFEI LVZHU BIOPHARM
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