Polypeptide vaccine production process

1 Proliferation and inactivation of NDV The NDV species were stored in sterile saline (containing penicillin, gentamicin) and diluted in a certain ratio for 15-20 min. Inoculation of 10 days old chicken embryos per 012 ml of embryos The allantoic cavity was incubated at 37 °C for 96 h, and the live embryo was cooled at 0 to 4 °C for 6-10 h. The chicken embryo allantoic fluid was aseptically collected into a sterilized large bottle, and 1 ml of the mixed allantoic fluid was aspirated to measure the HA titer. At the same time, the large amount of allantoic fluid in the large bottle was added to formalin at a ratio of 012%, and inactivated at 37 °C for 20 to 24 hours. After passing the sterility test, it was placed in a low-temperature freezer for storage.

2 Proliferation and inactivation of AIV The AIV strains preserved were diluted as described above, and 11-day-old chicken embryos were inoculated at 012 ml per embryo, and cultured at 37 °C for 30 hours to harvest the dead embryos and the urea solution of the viable embryos. The mixed allantoic fluid was used to detect the HA titer. At the same time, the allantoic fluid in the large bottle was added to formalin at a ratio of 013%, inactivated at 37 °C for 24 hours, and stored in a low temperature freezer after sterility test.

3 Oil phase preparation Take 94 parts of No. 7 white oil, and after mixing 6 parts of Si-80, add aluminum stearate in a certain ratio, then heat until completely dissolved to make oil phase for use.

4 Preparation of aqueous phase Take 94 parts of No. 7 white oil, and after mixing 6 parts of Si-80, add aluminum stearate in a certain ratio, then heat until completely dissolved to make oil phase for use.

Production process of two-oil emulsion inactivated vaccine:


1 The three kinds of virus-forming seedlings that have passed the test are mixed in a certain ratio, and 96 parts of the mixed virus solution are thoroughly mixed with 4 parts of the sterile Tween 80 to completely dissolve the Tween 80 to prepare an internal aqueous phase.

2 Mix a small amount of white oil with a certain amount of aluminum stearate, heat and melt, then mix with appropriate amount of 80 and white oil to make the oil phase, and make the final concentration of white oil, sibba 80 and aluminum stearate. A certain proportion was reached; the oil phase was autoclaved at 116 °C for 30 min and then used.

3 Appropriate amount of Tween 80, oleic acid and propylene glycol are thoroughly mixed to prepare an external aqueous phase emulsifier, which is then thoroughly mixed with the virus solution to prepare an external aqueous phase.

The vaccine was prepared by a two-step emulsification method.

The first step: according to the internal water phase and the oil ratio of 4:6, the internal water phase is slowly added to the low-speed stirring oil phase, after initial emulsification, high-speed emulsification for a few minutes, to make W / O oil Emulsion vaccine

Step 2: Add the W/O first-stage oil emulsion vaccine to the external aqueous phase according to the ratio of the W/O oil emulsion vaccine to the external aqueous phase at a volume ratio of 5:1, and emulsify at different emulsification rates. The vaccine was made into a triple emulsion vaccine against Newcastle disease infectious bronchitis and egg drop syndrome, and the stability of the vaccine prepared at different emulsification rates was examined.