Preparation Of Classical Swine Fever Virus Polyclone Antibodies And Their Binding With Pscudorabies Virus

Classical swine fever virus (CSFV) and pscudorabies virus (PRV) were proliferatedin pig passaged kidney cell line PK-15and baby hamster kidney passage BHK-21cells.After cultured for72hours, two viruses were harvested through cell disruption, salting out,dialysis and Sephadex G-75gel chromatography. Immunoaffinity chromatography columnwas prepared by coupling purified CSFV with epichlorohydrin activated Sepharose-4B asligands. This immunoaffinity chromatography column was used to purify antibodies fromthe serum of immunized pigs. The high immune rabbit serum was obtained by immuningthe rabbits using CSFV. Another purification method was that to use Q SepharoseTM-XLion exchange column to purify CSFV polyclonal antibodies from the high immune rabbitserum. The purity and activity of the obtained antibodies were detected by SDS-PAGE andindirect ELISA. The pig serum, the rabbit serum as well as the extracted antibodies fromthe pig serum and the rabbit serum, respectively, were detected using indirect ELISA toprobe their binding reaction with CSFV and PRV. The results showed that the purifiedCSFV antibodies by these two methods had higher purity, good activity and specificity.The purification methods used in this study were very rapid and feasible. The results ofcross-reaction showed that the CSFV antibodies not only reacted with CSFV, but alsobinded with PRV. The binding capacity of CSFV antibodies with PRV was approximatelyequivalent to half of that with CSFV. This study has certain guiding signification to thepreparation of therapeutic antibodies as well as the immunization work for classical swinefever and pscudorabies disease.