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Insect Cell Expression Systems
Protein Production Using Insect Cell Expression Systems
The insect cell expression and purification system is a versatile and robust platform used for producing a wide range of proteins. It offers distinct advantages, such as accurate post-translational modifications (PTMs), increased yields for secreted proteins, improved solubility, and the ability to support relatively high cell culture densities. These benefits make insect cell lines an excellent choice for expressing various types of proteins, including intracellular proteins, protein complexes, kinases, membrane proteins, and viral proteins.
At WuXi Biologics, our Protein Sciences (PS) Department provides a range of commonly used insect cell lines, such as Sf9 and HighFive. We leverage optimized expression vectors, host cells, and advanced virus packaging technology to ensure consistent and high levels of protein expression, resulting in the delivery of premium-quality proteins.
Insect Expression and Purification Services Workflow
Key Features of our Insect Expression and Purification Services:
- Use of the flashBAC™ system for accelerated timeline
- Small-scale expression condition optimization, including construct screening, MOI adjustment and harvest timing
- Advanced high-throughput purification system
- Scale up volumes up to 20 L
- Expertise with many types of proteins
Insect Expression and Purification Service Details:
| Service Item | Description | Duration | QC and Deliverable | Request A Quote |
| Baculovirus generation | P0, P1, P2 | 1-2 weeks | Titer: 1 x 109 PFU/mL | Request A Quote |
| 24 x 3 mL | 1 step purification, screening | 2-3 weeks | Purified protein | |
| 1 L – 20 L | 2-3 step purification | 4-5 weeks | SDS-PAGE, LC/MS | |
| Additional QC | RP-HPLC, bioburden, DSF, DSC, SPR, LC/MS, SEC-MALS, peptide mapping, cIEF, residual DNA, ELISA, Western blot, freeze thaw study, micro-developability testing | |||
Case Study: Using Insect Cell Expression Systems to Overcome Challenges in GPCR and Kinase Protein Expression and Purification
G-protein-coupled receptors (GPCRs) and kinases are two protein classes with significant therapeutic and research applications. However, expressing and purifying these proteins presents challenges related to stability, solubility, functionality, and low yield. In this case study, we demonstrate the use of our insect cell expression and purification services to achieve high-titer production of a GPCR protein and 8 kinase variants from Baculovirus pellets.
Figure A: We demonstrate high-titer production of a GPCR protein by optimizing the expression construct, expression, and purification processes. The achieved purity exceeded 90%, and the yield reached 1.5 mg/L. Additionally, we successfully stabilized the GPCR’s conformation using an appropriate ligand and effectively assembled GPCR-nanodiscs.
Figure B: Successful production of 8 kinase variants from Baculovirus pellets using an automated 2-step purification process. Despite differences in expression levels, all variants were expressed and purified in just 1 week, each reaching a purity of over 95%.
Frequently Asked Questions for Insect Expression and Purification Services
Q: What type of proteins has WuXi Biologics produced using insect expression systems?
A: Our Protein Sciences Department has extensive experience in producing a wide array of proteins using insect expression systems, including GPCRs, Ion channels, cell surface receptors, viral envelope proteins, kinases, proteases and enzymes, protein complexes, and other intracellular proteins.
Q: Between Sf9 and HighFive insect cells, which insect expression system do you use more often?
A: Sf9 cells are known for their superior protein expression levels compared to HighFive cells, and they are also easier to transfect. As a general practice, we begin with Sf9 cells due to these advantages. However, it’s worth noting that HighFive cells exhibit N-glycosylation patterns that resemble those of mammalian cells more closely. If the protein necessitates specific glycosylation patterns for correct folding or function, HighFive cells are the preferred choice.