Insect Cell Expression
The Baculovirus Expression Vector System (BEVS) is an established method within the scientific and industrial communities for generating large quantities of recombinant protein. BEVS is a eukaryotic system that utilizes the Autographa californica nuclear polyhedrosis virus (AcNPV) for high-level expression of recombinant proteins in Lepodoptera-derived insect cell lines, such as Sf9, Sf21, and High Five cells. Recombinant proteins produced using the system typically have correct folding, disulfide bond formation, and oligomerization, resulting in production of proteins that are functionally similar to their native forms. The system can accommodate large DNA inserts, perform intron/exon splicing, and yield relatively high levels of recombinant protein (0.1 to 50% of total protein). Short affinity tags, such as polyhistidine, Flag, and Twin-Strep, are often fused to recombinant proteins to aid in detection and purification of proteins. After affinity capture, tags can be removed using recombinant proteases such as TEV, HRV3C, thrombin, or SUMOStar.
Curia offers traditional and bacmid-based platforms for protein expression. Available platforms include Oxford Expression System flashBAC™, Bac-2-the-Future, and Thermo Fisher Bac-to-Bac®.
Curia’s Infection Kinetics Monitoring (IKM) technology was developed by our team of scientists to deliver unmatched reproducibility and scalability for baculovirus mediated expressions and this system is compatible with all popular kit systems that exist in the protein expression marketplace. IKM is a multi-component system that compiles a suite of real time cell population measurements, read during the course of a viral infection. In tandem with standard scouting measures, such as western blot, IKM substantially decreases the guesswork associated with baculovirus mediated scale-up resulting in unmatched scalability and reproducibility. This system also obviates the need for the laborious baculoviral titration saving our client’s time while making expression more predictable.
Baculovirus Infected Insect Cell (BIIC) is a method for preserving and scaling up baculovirus that provides a form of viral stock more stable than the traditional, extracellular stock. This powerful technology eliminates the need for virus amplification and re-titering, drastically reduces the turn-around time and resources required for scale-up and improves yield and consistency in protein expression. Curia offers this leading edge technology through one of the technology’s original co-inventors.
In addition, baculovirus particles (BVPs) are used to measure polyspecific binding of antibodies in an ELISA-based format for assessing therapeutic antibody developability.
|1036||Insect Scale-up Expression||Dependent on scale||Request|
|25610||Cloning into expression plasmid, and P1 virus generation||4-5 Weeks||Request|
|25620||30 ml small-scale expression analysis with affinity pulldown||5-7 Days||Request|
|25630||1 liter expression, and one-step affinity purification||5-7 Days||Request|
|25640||1 liter expression, and tandem affinity purification||6-8 Days||Request|
|25650||1 liter expression, and tandem affinity purification with SEC polishing||7-9 Days||Request|
|25670||Baculovirus Infected Insect Cell Stock Creation (BIIC)||5-7 Days||Request|
|25680||Small scale expression scouting and optimization||1-2 Weeks||Request|
|25690||Baculovirus particle (BVP) production for polyspecificity screening||2 Weeks||Request|