Patent ID: 9701946
Date: 2017-07-11
CPC Classifications: A61P,C12N

Claim:
1. A purification method for Embryo-derived avian Infectious Bronchitis Virus (IBV), comprising the following steps: 1) Inoculating fertilized eggs containing chicken embryos with avian Infectious Bronchitis Virus; 2) incubating the eggs containing the embryos at 37° C. for 48-96 hours, removing the eggs containing the avian embryos that have died within the first 24 hours during embryo incubation; refrigerating the remaining eggs containing the embryos at 2° C. to 8° C. for 8-16 hours for storage, then collecting the allantoic fluid from the eggs; 3) Centrifuging the allantoic fluid at 4000-6000 rpm for 10-50 min; 4) Ultrafiltrating the centrifugal liquid from step 3) with ultrafiltration membrane which pore size is 30-500 kDa, concentrating the filtrate to 20%-50% of the original volume; 5) Chromatographing the concentrated filtrate with a molecular sieve (with a column height of 30-120 cm packing); loading the concentrated filtrate with a loading volume (5% to 20% of the column volume, eluting the chromatography with PBS buffer solution, and collecting the first protein elution peak as avian infectious bronchitis virus; 6) Ultrafiltrating the avian infectious bronchitis virus elusion from step 5) with ultrafiltration membrane which pore size is 30-500 kDa and concentrating the filtrate to 20%-50% of the original volume.