Development of a Robust Protocol for the Production of Recombinant Proteins up to 20-L Scale Using BEVS

The aim of the study was to develop a scalable and robust generic process for production of recombinant cytosolic proteins in High Five cells using the baculo virus expression vector system (BEVS). The increasing demand for recombinant proteins in drug discovery has led to requirement for a generic protocol for protein supply. The process itself should fit into an ordinary weekly working scheme. The challenge was that the activity of many proteins could only be analysed when highly purified. Additionally, many proteins were extremely sensitive to protease degradation and could only be stored after purification. The influence of multiplicity of infection (MOI) and different growth media on protein expression was investigated. Time of infection and time of harvest was thoroughly evaluated with respect to high protein quality.