Preparation method of aluminum-containing adjuvant hepatitis B vaccine

A hepatitis B vaccine, aluminum adjuvant technology, applied in biochemical equipment and methods, medical preparations containing active ingredients, antiviral agents, etc., can solve the problem of high vaccination volume, low adjuvant adsorption rate, and antibody positive conversion rate Not ideal, etc., to achieve the effect of high antibody positive conversion rate, less adverse reactions, and improved immunogenicity

Active Publication Date: 2011-09-28
DALIAN HISSEN BIO-PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing aluminum adjuvant adsorption process for hepatitis B vaccine has problems such as low adjuvant adsorption rate, high vaccination volume and unsatisfactory antibody seroconversion rate.

Method used

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  • Preparation method of aluminum-containing adjuvant hepatitis B vaccine
  • Preparation method of aluminum-containing adjuvant hepatitis B vaccine
  • Preparation method of aluminum-containing adjuvant hepatitis B vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The preparation of embodiment 1 recombinant (hansenula) hepatitis B surface antigen stock solution

[0043] Fermentation: Take 1 strain of recombinant Hansenula working seed batch (obtained from the original strain of recombinant Hansenula hepatitis B vaccine with strain number HBsAgU35-16-9), inoculate in 300ml culture medium After culturing at 35°C for 24 hours, transfer to 3L medium, culture at 35°C for 24 hours, then transfer to 30L medium, culture at 35°C for 15 hours, and finally transfer to 200L medium, culture at 35°C, control The dissolved oxygen is greater than 20%, the pH is 6.8, and the air flow rate is 30-200ml / h. After culturing for 65 hours, collect about 240L of Hansenula cell fermentation liquid expressing the surface antigen of hepatitis B virus.

[0044] Preliminary purification: Grinding and crushing Hansenula cell fermentation liquid by physical crushing method to achieve a crushing rate of 92%, centrifuging at a speed of 4000rpm to remove cell debr...

Embodiment 2

[0046] Embodiment 2 preparation of hepatitis B vaccine semi-finished product (aluminum adjuvant+HBsAg technology-prior art), semi-finished product batch number is S201001

[0047] Aluminum adjuvant preparation: start stirring, add 10% AlCl to the reaction bottle 3 Solution 54ml, add 0.5mol / L NaOH solution at a speed of 50ml / min, take samples at any time to measure the pH value, when the pH value reaches 7.00, stop adding the solution, add 110ml of 0.5mol / L NaOH solution, and then add 0.9% NaCl solution 436ml with a final volume of 600ml, and then sterilized by moist heat at 121°C for 30 minutes to be ready-to-use aluminum adjuvant.

[0048] Dilution of the stock solution: 54.5ml of the stock solution (prepared by Example 1) with a protein content of 220 μg / ml was placed in a 500ml Erlenmeyer flask, and 300ml of 0.9% NaCl solution was added to make the protein content 40 μg / ml.

[0049] Adsorption of semi-finished product: Start stirring, add 300ml of aluminum adjuvant to anot...

Embodiment 3

[0050] Embodiment 3 preparation of hepatitis B vaccine semi-finished product (aluminum adjuvant+HBsAg technology-prior art), semi-finished product batch number is S201002

[0051] Aluminum adjuvant preparation: start stirring, add 10% AlCl to the reaction bottle 3 Solution 54ml, add 0.5mol / L NaOH solution at a speed of 50ml / min, take samples at any time to measure the pH value, when the pH value reaches 6.95, stop adding the solution, add 0.5mol / L NaOH solution 108ml, and then add 0.9% NaCl solution 438ml with a final volume of 600ml, and then sterilized by moist heat at 121°C for 30 minutes to prepare aluminum adjuvant for use.

[0052] Dilution of the stock solution: 54.5ml of the stock solution (prepared by Example 1) with a protein content of 220 μg / ml was placed in a 500ml Erlenmeyer flask, and 300ml of 0.9% NaCl solution was added to make the protein content 40 μg / ml.

[0053] Adsorption of semi-finished product: Turn on stirring, add 300ml of aluminum adjuvant to anoth...

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Abstract

The invention discloses a preparation method of an aluminum-containing adjuvant hepatitis B vaccine, belonging to the biotechnology field. The preparation method is characterized in that an aluminum adjuvant Al(OH)3 is produced by an on-line reaction, i.e. after a phosphate buffer solution (PBS), a KAl(SO4)2 solution and a hepatitis B surface antigen stock solution are mixed, an NaOH solution is added to the mixed solution, an Al(OH)3 adjuvant is continuously produced, and simultaneously, hepatitis B surface antigens are continuously coated and adsorbed; and the process is called 'in-situ adsorption'. In the invention, the Al(OH)3 adjuvant is produced by an in-situ reaction to greatly improve the adsorption rate of the hepatitis B surface antigens, thereby improving the immunogenicity of the antigens, being capable of more effectively causing organisms to generate an immune response, and producing more protective antibodies. The practice proves that the aluminum adjuvant hepatitis B vaccine produced by the method disclosed by the invention has the advantages of small inoculation amount, few adverse responses, high antibody positive conversion rate and the like, and can induce high-level antibody response after being immunized. Simultaneously, the processing steps are also simplified, and the production cost is greatly lowered.

Description

technical field [0001] The invention relates to a preparation method of a vaccine, in particular to a preparation method of an aluminum-containing adjuvant hepatitis B vaccine, and belongs to the field of biotechnology. Background technique [0002] There are about 350 million people infected with hepatitis B virus (HBV) in the world at present, and there are 130 million people with chronic HBV infection in my country. Chronic HBV infection is a serious threat to human health, persistent infection will develop into chronic hepatitis, liver cirrhosis, liver cancer and so on. At present, there is no cure for hepatitis B in the medical field. Large-scale vaccination of hepatitis B vaccine is the most economical and effective way to prevent HBV infection, and it is also the fundamental measure to reduce the harm of HBV. [0003] The main component of hepatitis B vaccine is hepatitis B surface antigen (HbsAg), and the adjuvant is traditional aluminum hydroxide. [0004] In 1926...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P31/20A61K39/39A61P1/16A61K39/29
CPCA61K39/39A61K39/29A61K2039/55505A61K39/292C12N2730/10134A61K39/12A61P1/16A61P31/20
Inventor 李贵兴张平张海春陈新亭李晓宇张娟
Owner DALIAN HISSEN BIO-PHARM CO LTD
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