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Rabies immunogenic conjugate

A technology of immunogenicity and conjugates, applied in the medical field, can solve the problems of highly dangerous parts, inability to produce effective protection, backwardness, etc., and achieve the effect of enhancing immunogenicity

Active Publication Date: 2018-06-19
复星安特金(成都)生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] It can be seen that the existing inactivated whole virus particle vaccines are based on the fatal defects of no or weak cellular immunity and slow humoral immunity, and the production of antibodies obviously lags behind the speed and speed at which rabies virus reproduces in local muscles and invades peripheral nerves. Time, especially for those exposed with severe bites, highly dangerous parts, and short incubation period, cannot produce effective protection (Wang Shusheng. Applied Preventive Medicine. 2010.16(1):1‐4)

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0229] Rabies whole virus particle antigen stock solution preparation and vaccine preparation:

[0230] Cells from the Vero cell working bank were recovered and expanded at 37°C, and after 3-5 generations of continuous passage, the cell density reached 1.0×10 5 ~1.0×10 6 / ml, inoculated into a bioreactor containing 1-25g / L microcarriers for tank-flow culture. Microcarrier high-density suspension culture for 5 to 7 days, the cell density reaches 1.0×10 6 ~1.0×10 7 / ml, replace with maintenance solution. Inoculate the virus titer at 0.001~0.1MOI not less than 7.5lg LD 50 / ml of rabies virus fixed virus CTN‐1V strain. Replace with maintenance medium, and culture in tank flow at 33-35°C. The virus fluid can be continuously harvested on the third day, and the virus fluid can be continuously harvested for more than 20 days. The harvested virus fluid is clarified and filtered, and concentrated by 100-300KD membrane ultrafiltration for 10-30 times. Add β‐propiolactone at a rat...

Embodiment 2

[0233] Preparation of carrier protein‐tetanus toxoid (TT)

[0234] Open the tetanus freeze-dried bacterial strain preserved at low temperature, inoculate it into a semi-solid medium containing tryptone beef infusion agar, and cultivate it at 33-37°C for 24-48 hours. Transfer to semi-solid medium, and cultivate at 33-37°C for 24-48 hours. Then transfer to the strain bottle containing tryptone beef extract liquid medium to carry out enrichment culture, and cultivate at 33-37 DEG C for 24-48 hours. The culture in the strain bottle is transferred to a fermenter of double peptone liquid medium, and cultured at 33-37° C. for 60-80 hours. After the fermentation is completed, formaldehyde with a final concentration of 0.1-0.5% is added for sterilization treatment, and the supernatant is collected by centrifugation. The supernatant was clarified and filtered through a 0.45-1.2 μm filter, and then concentrated by a 50-100KD membrane ultrafiltration. Add NaHCO separately 3 The final ...

Embodiment 3

[0236] Rabies Whole Virus Particle-Carrier Protein Immunogenic Conjugate Preparation and Vaccine Preparation:

[0237] Measure the stock solution of tetanus toxoid and dilute it to 1-10 mg / ml with 0.1 M PBS with pH 7.5. Add 100mg / ml CDAP (1-cyano-4-dimethylamino-pyridine tetrafluoroboric acid) acetonitrile solution to a final concentration of 0.5-1.5mg / mgTT, react for 30 seconds, add 0.2M TEA (triethylamine), The ratio of TEA addition to CDAP addition is 1-3:1 (v / v), adjust the pH to 9.0, and maintain the reaction for 2-5 minutes. Add ADH (adipic hydrazide) NaHCO 3 (0.1M) solution to a final concentration of 2.0-4.0 mg / mgTT, and maintain the reaction for 120 minutes. Use 50-100KD membrane ultrafiltration to remove residual chemical reagents, which is the stock solution of tetanus toxoid‐ADH derivatives. Measure the purified stock solution of rabies whole virus particles prepared according to the method of Example 1 and without adding human serum albumin, according to the ra...

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Abstract

The invention relates to a rabies immunogenic conjugate (RIC). The rabies immunogenic conjugate comprises a rabies virus antigen (RabAg) and carrier protein (CP), and is characterized in that the rabies virus antigen at least comprises the rabies virus antigen connected with the carrier protein through a chemical bond; the carrier protein at least comprises the carrier protein connected with the rabies virus antigen through the chemical bond; the chemical bond connects the rabies virus antigen and the carrier protein to form a rabies virus antigen-carrier protein immunogenic conjugate. The invention further relates to a novel rabies conjugate vaccine (RCV) prepared according to the rabies immunogenic conjugate and a preparation method thereof. The invention preliminarily proves that the rabies immunogenic conjugate overcomes the defects in the prior art and has a relatively good effect.

Description

[0001] This application is a divisional application with the application number 201710381237.2, the application date is May 25, 2017, and the invention title is "a rabies immunogenic conjugate and vaccine and its preparation method". technical field [0002] The invention relates to the medical field, in particular to vaccines and vaccine preparations, in particular to a rabies immunogenic conjugate and a preparation method thereof, and a rabies conjugate vaccine and a preparation method thereof. Background technique [0003] Rabies (Rabies) is an acute infectious disease caused by Rabies Virus (Rabies Virus) infection, and is a zoonotic acute infectious disease of natural foci. The rabies virus is mainly transmitted to humans through direct contact with damaged skin or mucous membranes through the saliva of infected animals. Once rabies occurs, its case fatality rate is almost 100%. Dogs are the main host of rabies virus, and up to 99% of human rabies is transmitted by dog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/205A61K39/385A61P31/14
CPCA61K39/12A61K39/385A61K2039/5258A61K2039/6031A61K2039/6037C12N2760/20134
Inventor 薛平李银波王超赵光伟王岩杨冬妮
Owner 复星安特金(成都)生物制药有限公司
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