Study On The Multi-step Continuous Integrated Purification Method Of Foot And Mouth Disease Virus Antigens

In this thesis,a multi-step purification method for the inactivated virus of the popular strains of foot-and-mouth disease A and O was carried out,including high-speed centrifugation,tangential flow ultrafiltration concentration,PEG6000 purification,and ion exchange chromatography and hydrophobic chromatography.High-speed centrifugation can remove cell debris and reduce the turbidity of the feed solution.After centrifugation,the concentration of FMDV O-type 146 S is 3.15 μg/m L,the protein content is 6560 μg/m L,the A-type 146 S concentration is 4.2 μg/m L,and the protein content is 7520 μg/m L;Depth filtration can further remove large particles of impurities and reduce turbidity.The recovery rate of FMDV O-type antigen is 89.96 %,and the recovery rate of A-type antigen is 89.87 %;TFF ultrafiltration can be highly concentrated,reduce the volume of feed liquid and remove some impurities,FMDV The recovery rate of O-type antigen is 87.94 %,and the recovery rate of A-type antigen is 94.80 %;PEG6000 purification can further reduce the volume of feed solution and replace the buffer.After purification,the recovery rate of FMDV O-type antigen is 82.13 %,and the purity of O-type antigen is 16 %,the recovery rate of A-type antigen was 84.24 %,the purity of A-type antigen was 15 %;the recovery rate of FMDV Otype antigen after ion exchange chromatography was 83.64 %,the purity of O-type antigen was 58 %,and the recovery rate of A-type antigen was 94.63 %.The purity of type A antigen was 56 %;the recovery rate of FMDV type O antigen was 95.44 % after hydrophobic chromatography,the purity of type O antigen was 85 %,the recovery rate of type A antigen was 90.20 %,and the purity of type A antigen was 81 %.Through the multi-step purification method in this experiment,the final concentration of FMDV type O antigen can reach 208 times,the antigen recovery rate is 51.87 %,the concentration of 146 S is 340.38 μg/m L,and the purity is 85 %;the final concentration of FMDV type A antigen can reach 143 times.times,the recovery rate is 61.25 %,the concentration of 146 S is 367.53 μg/m L,and the purity is 81 %,which fully meets the expected effect.In the case of high immune effect of animals,it plays a good role.prevention and control effect.