Isolation,Purification and Immunogenicity Evaluation of Senecavirus A Intact Particle and Empty Capsid
Senecavirus A(SVA)has the ability to form intact virions and empty capsids during replication;however,the conditions for isolating them and their differences in antibody response are still unknown.In this study,we utilized guanidine hydrochloride to generate SVA empty cap-sids and determined the optimal conditions for separating SVA intact virions and empty capsids.This was achieved through ultracentrifugation using different density gradients,media,rotation rates,and durations.Subsequently,SVA intact virions and empty capsids were intramuscularly immunized into BALB/c mice at a dosage of 12.5 µg per mouse.Specific and neutralizing anti-bodies were then monitored 1-7 weeks post-immunization.The findings indicated that the addi-tion of 100 mmol·L-1 guanidine HC1 for 2 hours in infected cells at MOI=1 of SVA resulted in the complete formation of empty capsids.The most effective method for distinguishing SVA in-tact virions from empty capsids was centrifugation at 36 000 r·min-1 for 2.5 hours,using a cesi-um chloride gradient ranging from 10％to 50％.Furthermore,the specific and neutralizing anti-body levels for SVA intact virions were comparable to those of empty capsids.This study pro-vides a new reference for the isolation and purification of the complete and empty capsid of Seneca virus A,as well as for the development of recombinant SVA virus-like particle vaccines.
万方数据
维普
