Preparation method of hepatitis vaccine and its use

A hepatitis B vaccine and a new type of technology, applied in the field of human hepatitis B vaccine and its preparation, can solve the problems of granulomatous inflammation, contact allergic reaction, local reaction, etc., achieve reduction of inflammatory nodules, reduction of inflammatory response, The effect of reducing side effects

Inactive Publication Date: 2006-08-30
THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] ①Long use time: It has been used since 1926; ②Safe and effective; ③Inexpensive; ④Widely used; ⑤Weak adjuvant effect or no adjuvant activity for some vaccines; ⑥Easy to induce the expression of IgE; Erythema, subcutaneous nodules, contact allergic reactions, granulomatous inflammation, etc., and some even have serious local reactions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Preparation method of nano aluminum adjuvant involved in the present invention

[0033] Take a certain amount of AlCl3, cyclohexane and distilled water in a beaker, then add the benzalkonium bromide-n-octanol-cyclohexane-AlCl3 solution system, and then stir with a high-speed dispersing homogenizer to make it emulsified into micro Lotion. Pour into a reaction flask with a lid, heat and stir, slowly add excess ammonia water dropwise, close the lid, and react for 3 hours, which is Al(OH)3 sol. Use freeze drying or vacuum drying. Then it is dissolved in water and dispersed by ultrasonic, and it is dispersed in the aqueous solution. The method of electron microscope observation and measurement to determine the nanoparticle diameter.

Embodiment 2

[0034] Example 2. Preparation method of new type human hepatitis B vaccine

[0035] The prepared nano-aluminum adjuvant is made up to 0.1 mg per milliliter (using water as a solvent), the nano-aluminum adjuvant solution is dispersed by ultrasound for 20 minutes, and the commercially available non-adjuvant hepatitis B vaccine for humans is added at a dose of 5.0 micrograms per tube. Rabies vaccine and nano aluminum adjuvant were mixed in a volume of 3:1, adsorbed at 4°C for one week, centrifuged at 15,000 rpm for 30 minutes, and the concentration of rabies vaccine antigen in the supernatant was detected by ELISA double antibody sandwich method. According to the method introduced above, the amount of the new human hepatitis B vaccine adsorbed on the unit mass of nano aluminum adjuvant is 10-20 times higher than that of the conventional aluminum adjuvant.

Embodiment 3

[0036] Example 3. Results of mice immunized with the novel hepatitis B vaccine of the present invention

[0037] Main reagents and their preparation

[0038] The composition of the coating diluent (0.05mol / L sodium carbonate-sodium bicarbonate buffer, pH 9.6):

[0039] Na2CO3 1.5g, NaHCO3 2.9g, Na2N3 0.2g, add double distilled water to 1000ml, adjust to pH 9.6.

[0040] Composition of blocking solution (5% calf serum / PBS solution):

[0041] 50ml of calf serum was added to 950ml of PBS (pH7.4).

[0042] Phosphate buffer (PB)

[0043] A solution (0.2mol / L sodium dihydrogen phosphate aqueous solution) composition: NaH2PO4·H2O 27.6g dissolved in 1000ml ultrapure water.

[0044] B solution (0.2mol / L disodium hydrogen phosphate aqueous solution) composition: Na2HPO4·7H2O 53.6g (or Na2HPO4·12H2O 71.6g or Na2HPO4·2H2O 35.6g) dissolved in 1000ml of ultrapure water.

[0045] Sample dilution solution (PBS, 0.01mol / L phosphate buffered saline) components: 19ml PB A solution; 81ml PB B solutio...

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Abstract

A human rabies vaccine is prepared from the existing hepatitis B vaccine without adjuvant through adsorbing it by the aluminum hydroxide nanoparticles. Its adsorbed capacity is increased by 10-20 time. It can be used for emergency inoculation for preventing hepatitis B as it has the fast release effect.

Description

Technical field [0001] The invention relates to a hepatitis B vaccine for human use and a preparation method thereof, in particular to a hepatitis B vaccine containing a nano aluminum adjuvant and a preparation method thereof. technical background [0002] The main component of hepatitis B vaccine is HBsAg with an isoelectric point of 4.5. HBsAg contains a large number of phosphate groups. Because the isoelectric points (iep) of HBsAg and aluminum hydroxide adjuvant are 4.5 and 11.4, respectively. Therefore, in the neutral environment of the body with pH=7.4, the former shows a negative charge and the latter shows a positive charge. The two can have a good adsorption effect due to electrostatic force. [0003] The main mechanisms of aluminum adjuvant adsorption of antigens include electrostatic force, hydrophobic interaction and group exchange [Vogel FR, Hem SL. In: Plotkin SA, Orenstein MD, editors. Vaccines. 4th ed. New York: Sanders]. The adsorption caused by hydrophobic inter...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/29A61K9/14A61P31/20A61P1/16
Inventor 吕凤林何萍胡承香
Owner THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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