Abstract
Among the wide range of methods and expression hosts available for the heterologous production of recombinant proteins, insect cells are ideal for the production of complex proteins requiring extensive post-translational modification. This review article provides an overview of the available insect-cell expression systems and their properties, focusing on the widely-used Baculovirus Expression Vector System (BEVS). We discuss the different strategies used to generate recombinant baculovirus vectors and show how advanced techniques for virus titer determination can accelerate the production of recombinant proteins. The stable transfection of insect cells is an alternative to BEVS which has recently been augmented with recombinase-mediated cassette exchange for site-specific gene integration. We consider the advantages and limitations of these techniques for the production of recombinant proteins in insect cells and compare them to other expression platforms.
Highlights
Many different expression systems are available for the production of recombinant proteins, each with numerous options
The baculovirus expression vector system is often used with cell lines Sf-9 and Sf-21, both originating from cell line IPLB-SF-21 isolated from Spodoptera frugiperda pupal ovarian tissue (Vaughn et al, 1977), as well as BTI-TN-5B1-4, established from ovarian cells of the cabbage looper Trichoplusia ni (Granados et al, 1994) and better known under the brand name High FiveTM
The baculovirus expression system is generally superior because the strong very late promoters achieve high levels of expression. This does not apply to every protein, e.g., immediate early promoters in stably-transfected cells may be more efficient for the expression of membrane-bound or secreted proteins, because processing may already be compromised during the late stages of baculovirus infection (Jarvis et al, 1990; 1996)
Summary
Many different expression systems are available for the production of recombinant proteins, each with numerous options. The production of a recombinant protein is usually motivated by an ambition to determine the protein structure, investigate its activity or search for interaction partners in order to unravel its mode of action. The simplest or most accessible system that meets minimum requirements is often chosen for initial synthesis than after heterologous expression, because the starting material is less complex. Chemical synthesis reaches its limits when the peptide exceeds approximately 70 amino acids in length, contains a high proportion of challenging amino acids (e.g., arginine, cysteine, methionine and tryptophan) or requires post-translational modification. Peptides containing multiple disulfide bonds can be synthesized (Reinwarth et al, 2012) success depends on chance and laborious procedures are required to verify the disulfide linkages. Expression studies and if successful these efforts are scaled up for downstream applications
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
