Baculovirus-Insect Cell Expression

At Creative BioMart, our team of experts have developed baculovirus-insect cell expression platform for virus production and recombinant protein manufacture in various scales. The ability of Creative BioMart's baculovirus-insect cell system to express proteins with complex post-translational modifications (PTMs), combining with high production yield, makes it perfectly suitable for generating mammalian proteins, especially cellular signaling proteins including protein kinases, protein phosphatases and cellular protein substrates, amongst others.

What is Baculovirus-Insect Cell Expression System?

The baculovirus-insect cell expression system is a binary system consisting of two basic components, including a baculovirus expression vector that delivers a foreign gene encoding a protein of interest to the host and its host, an insect cell line, usually a lepidopteran insect cell line.

The baculovirus-insect cell expression system utilizes recombinant baculoviruses (insect viruses) and their ability to manufacture high yields of biologically active proteins from infected insect cells. The proteins of interest can be easily purified from infected cells or their supernatants using tag and affinity chromatography. The important steps of the baculovirus-insect cell expression system include baculovirus production, insect cell infection and purification.

Advantages of Baculovirus-Insect Cell Expression System

• Improved solubility
• Able to perform complex PTMs
• High protein expression levels compared to other higher eukaryotes
• Suitable for the production of large proteins and protein complexes
• Capable of producing cytotoxic proteins
• Easy to scale up in bioreactors
• Lower-cost
• Operational safety: baculovirus are noninfectious to vertebrates

Various Methods of Generating Recombinant Baculovirus Systems

In order to adapt to the different purposes of customers, Creative BioMart offers various approaches to generate a recombinant baculovirus system containing the gene of interest.

System	Features
Bac-to-Bac	Requires integration with baculovirus shuttle vector (Bacmid) in bacterial host cell before transfection.
flashBAC	Transfection with linear DNA, recombination occurs within host insect cells. Deletion of orf1629 enables better secretion of recombinant protein.
BaculoGOLD	Transfection with linear DNA, recombination occurs within host insect cells.
BaculoDirect	Pre-linearized DNA. Requires Ganciclovir in culture.
BestBac	Transfection with linear DNA, enhance production of target protein.

Service Content of Baculovirus-Insect Cell Expression System

1

Gene Synthesis Plus Codon Optimization

• Leading codon optimization system
• Analysis and optimization of rare codons, codon bias and other factors

Timeline: 1-2 weeks

2

Clone Target Gene into Expression Vector

• Plasmid sequencing
• Plasmid preparation
• Generation of recombinant Bacmid

Timeline: 1 week

3

Transfect Baculovirus with Transfer Vector

• Virus packaging
• Virus amplification

Timeline: <1 week

4

Insect Cells Infection with Baculovirus

• Multiple insect cell lines
• Infection of insect cells with P1 or P2 generation viruses

Timeline: <1 weeks

5

Protein Expression and Purification

• Small-scale fermentation culture
• Expression condition optimization
• Protein expression evaluation
• Various protein purification methods

Timeline: 2-4 weeks

6

Optional Services

• Secondary purification
• Tag removal
• Endotoxin removal
• Protein labeling
• Activity assay
• Protein analysis
• Other more

Timeline: 1-2 weeks

7

Quality Control

• SDS-PAGE
• Western blot
• Mass spectrometry

Timeline: <1 week

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Deliverables

• Purified protein [supplied in liquid or lyophilized form]
• Gene sequencing report
• QC report
• SDS-PAGE/ HPLC/ SEC
• Activity data if applicable

Insect Cell Line Options from Creative BioMart

Insect Cell Line	Source	Application
Sf9 and Sf21	Spodoptera frugiperda	All general types of recombinant protein expression.
MimicTM Sf9	Spodoptera frugiperda	Expression of mammalian glycoproteins.
High FiveTM	Trichoplusia ni	Secretion of recombinant proteins. Shorter culture period.

Reference:

1. Unger T and Peleg Y. Recombinant Protein Expression in the Baculovirus-Infected Insect Cell System. Methods Mol Biol. 2012;800:187-99. doi: 10.1007/978-1-61779-349-3_13.