Transforming lepidopteran insect cells for improved protein processing and expression

Abstract

The lepidopteran insect cells used with the baculovirus expression vector system (BEVS) are capable of synthesizing and accurately processing foreign proteins. However, proteins expressed in baculovirusinfected cells often fail to be completely processed, or are not processed in a manner that meets a researcher’s needs. This chapter discusses a metabolic engineering approach that addresses this problem. Basically, this approach involves the addition of new or enhancement of existing protein processing functions in established lepidopteran insect cell lines. In addition to improvements in protein processing, this approach has also been used to improve protein expression levels obtained with the BEVS. Methods for engineering cell lines and assessing their properties as improved hosts for the BEVS are detailed. Examples of lepidopteran insect cell lines engineered for improved protein N -glycosylation, folding/trafficking, and expression are described in detail.