Production and Purification of Yellow Fever Vaccine using Chick Embryonic Fibroblast Cells

Abstract

Background newlineYellow fever is a viral haemorrhagic fever caused by the Yellow Fever Virus, which belongs to the Flaviviridae family and Flavivirus genus. This small (50 nm) virus has a nucleocapsid with a 13 kDa core protein, enclosing RNA that is single-stranded and positive-sense RNA. Various vaccine manufacturers presently produce the yellow fever vaccine using embryonated eggs, method that has remained largely unchanged from 1930s. WHO reports indicate a yellow fever mortality rate of 20 50%. newlineAim newlineThe purpose of this research is to examine the production of the yellow fever vaccine utilizing the 17D strain, while also enhancing its titre through the statistical optimization of process parameters and the purification of the yellow fever virus. Additionally, this study seeks to optimize the most effective stabilizer for the formulation of the YF virus. newlineMaterials and Methods newlineThe SPF eggs were sourced from Venky Chicken Pvt Ltd in Pune, India, while the live- attenuated yellow fever virus 17DD was provided from the WHO (World Health Organization) for the optimization studies. A custom design approach was utilized to optimize the placement of the SPF egg, relative humidity, and incubation duration. A Central Composite Design (CCD) was applied to ascertain the optimal volume of trypsin and incubation temperature. Additionally, a Mixed Design was employed to assess various media, including DMEM (M1), EMEM (M2), VP-SFM AGT (M3), VP-SFM (M4), OptiPRO-SFM (M5), EX-CELL-293 (M6), VeroVax (M7), OptiVERO (M8), VCum- Lsx0032 (M9), M10-CDM4Avian media (M10), HEK293 (M11), and CDM4HEK293 (M12). The yellow fever virus was purified through Tangential Flow Filtration (TFF), affinity chromatography using cellufine sulfate, diafiltration for buffer exchange, and subsequent microfiltration. concentrations of sucrose and gelatin were tested as adjuvants in the formulation of the yellow fever drug product. newline newline