3.3. Production, purification, and characterization of EV71 VLPs
Fig. 3. Production, purification, and characterization of EV71 VLPs. For EV71 VLP production by ΔM51 vector, BHK-21 CHIR-98014 were co-infected with mP1:m3CD = 9:1 (mP1/m3CD) or with only mP1-3CD at a total MOI of 0.01. The viruses were concentrated via ultracentrifugation and then purified through discontinuous CsCl gradient. Seven fractions (based on the bands in the ultracentrifuge tube) were taken from top to bottom and analyzed via Western blotting (A, B, D, E) and TEM examination (C, Image may be NSFW.
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F). (A), (B) and (C): the VLP produced by mP1/m3CD co-infection; (D), (E) and (F): the VLP produced by mP1-3CD. To gain EV71 natural virions, RD cells were infected with EV71 (genotype C4) at a total MOI of 0.1, the viruses were ultracentrifuged through 20% sucrose. The pellet was resuspended in PBS for characterization via TEM examination (C).Figure optionsDownload full-size imageDownload as PowerPoint slide
3.4. Attenuation of rVSVs in vivo
Fig. 3. Production, purification, and characterization of EV71 VLPs. For EV71 VLP production by ΔM51 vector, BHK-21 CHIR-98014 were co-infected with mP1:m3CD = 9:1 (mP1/m3CD) or with only mP1-3CD at a total MOI of 0.01. The viruses were concentrated via ultracentrifugation and then purified through discontinuous CsCl gradient. Seven fractions (based on the bands in the ultracentrifuge tube) were taken from top to bottom and analyzed via Western blotting (A, B, D, E) and TEM examination (C, Image may be NSFW.
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F). (A), (B) and (C): the VLP produced by mP1/m3CD co-infection; (D), (E) and (F): the VLP produced by mP1-3CD. To gain EV71 natural virions, RD cells were infected with EV71 (genotype C4) at a total MOI of 0.1, the viruses were ultracentrifuged through 20% sucrose. The pellet was resuspended in PBS for characterization via TEM examination (C).Figure optionsDownload full-size imageDownload as PowerPoint slide3.4. Attenuation of rVSVs in vivo
