Domestic animal and aquatic product causal agent resistant specific IgY or compound IgY and application thereof
A specific, pathogenic technology, applied in the direction of antibodies, anti-infective drugs, drug combinations, etc., can solve the problems of low activity, low antibody activity, and doubtful effects.
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Embodiment 1
[0172] Example 1: Preparation of anti-porcine transmissible gastroenteritis and epidemic diarrhea virus-specific compound duck IgY by genetic engineering
[0173] 1. Prepare the expressed protein antigen components of the two viruses, specifically comprising the following steps:
[0174] The signal peptide and the fragments of the transmembrane region were removed from RNA cloned genes resistant to porcine transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV) respectively by RT-PCR method. Insert the pGEM-T trimmer first. After sequencing proved that the obtained gene sequence was correct, it was digested with restriction endonucleases EcoR I and Not I, and the target fragment was recovered by electrophoresis, and connected with the yeast expression vector pPIC9K digested with the same double enzymes. Transform E. coli. Positive clones were picked, plasmids were extracted, and after enzyme digestion and identification were correct, Pichia past...
Embodiment 2
[0186] Example 2: Using fish vaccines as antigens to prepare anti-aquatic common virus-specific turkey IgY
[0187] The specific implementation steps are as follows:
[0188] Purchase 50 ready-made grass carp haemorrhagic disease vaccines (anti-reovirus) and channel catfish virus vaccines (300μg / 1.5mL each) from the Fisheries Research Institute of Fujian Province in China. The virus components are separated in the machine. Add sodium dodecyl sulfate (SDS) to the final concentration of 1.8-2.5%, and lyse at 37° C. for 30 minutes to obtain a virus lysate.
[0189] Then add this lysate into Freund's adjuvant at a ratio of 1-10:10-1, put it into a high-speed homogenizer, and homogenize at a high speed of 30,000rpm to form a water-in-oil liquid, which is to make a vaccine component and split antigen , 150mL in total. Then immunize 50 healthy egg-laying turkeys with this vaccine-type antigen, inject 1 mL intramuscularly into each chicken, and inject once every two weeks, a total ...
Embodiment 3
[0195] Example 3: The nanometerization of anti-porcine transmissible gastroenteritis and epidemic diarrhea specific compound IgY and anti-bovine transmissible gastroenteritis and epidemic diarrhea specific compound IgY improves bioavailability and further enhances its curative effect
[0196] Preparation of nano-IgY dry powder:
[0197] The anti-porcine transmissible gastroenteritis and epidemic diarrhea specific compound IgY and the anti-bovine transmissible gastroenteritis and epidemic diarrhea specific compound IgY dry powder prepared by the present invention are processed into a particle size between 1 and 100 nm by ultra-fine method. Ultrafine particles become nanoscale IgY ultrafine powder. In the present invention, one of the two IgYs is added into an "ultrafine pulverizer" for grinding and crushing to obtain ultrafine particles with a mesh size of ≥15,000. The anti-porcine transmissible gastroenteritis and epidemic diarrhea specific compound IgY and the anti-bovine tr...
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