Table 1 Generation and characterization of ZIKV sub-isolates used to derive PIZV candidates.

From: Purified Inactivated Zika Vaccine Candidates Afford Protection against Lethal Challenge in Mice

 

Virus stock generation

Characterization

CDC

Virus isolate amplified in Vero (2×) and C6/36 (1×)(V2/C6-1)

Plaque titer/phenotype, identity by RT-PCR, mycoplasma and togavirus testing

P1

Virus amplification in Vero

TCID50 titer

P2

Plaque purification of P1

plaque phenotype

P3

Plaque purification of P2

plaque phenotype

P4

Plaque purification of P3

plaque phenotype

P5

Amplification of P4 plaques (a-f sub-isolates) in Vero

TCID50 titer

P6

Amplification of P5 virus (a-f sub-isolates) in Vero

TCID50 titer, plaque phenotype, full genome sequencing, growth kinetics in Vero

P7

Amplification of P6 (b and e sub-isolates) in Vero; Used for deriving PIZV

Relative purity (SDS-PAGE), antigenicity (immunoblot, ELISA), EM, completeness of inactivation (serial cell culture passaging)