Insect Cell Protein Expression Services

  

 

 

 

Insect cells and baculovirus expression has been a powerful protein expression system for intracellular protein production. It also works very well for some viral proteins that may pose negative effects on mammalian cells. Insect cells can grow to very high cell density and can achieve very high protein yield after infected with baculovirus containing the target gene of interest.

KMD Bioscience has extensive experiences in producing large quantities of recombinant proteins with insect expression system. We are particularly good at purifying intracellular proteins from insect cells. In the past couple of years, we have successfully produced about 1000 recombinant proteins using our own insect expression systems. Many of these proteins are difficult-to-express proteins, including many kinase proteins, viral proteins, and some cytokines and growth factor proteins.

 

KMD Bioscience can provide the following insect host cell lines:

Cells

Source

Applications

Sf9

Sf9 cells are derived from the IPLBSF-21 cell line.

Transfection, purification, production of high titer viruses, plating and expression of recombinant proteins. Sf21 cells are superior to Sf9 cells in some structural expression.

Sf21

Sf21 is a late embryonic cell of drosophila. Most of them are female tetraploid cells and some are diploid cells.

More suitable for recombinant expression of viral structural proteins.

Hi5

Hi5 is a late embryonic cell of drosophila. Most of them are female tetraploid cells and some are diploid cells.

More suitable for recombinant expression of viral structural proteins.

S2

S2 is a late embryonic cell of drosophila. Most of them are female tetraploid cells and some are diploid cells.

More suitable for recombinant expression of viral structural proteins.

 

Insect Protein Expression Services Procedure:

 

Steps

Specification

Timeline

Deliverables

Subcloning

* Gene synthesis and codon optimization
* Subcloning into expression vector

2 weeks

* Construct report

Culture expansion of virus

* Titer test108-109) and verification of protein expression
* P3 virus production (Optional)

3-4 weeks

* 5-10 ml, >107 pfu/ml recombinant baculovirus
* Experiment report

Screening of yeast expression strains

* Detection of recombinant protein expression

2 weeks

* Positive clone strains
* Screening report

Expression optimization and testing

* P3 virus transfected insect cells
* SDS-PAGE &WB

1-2 weeks

* Images
* Experiment report

Large-scale protein expression and purification

* The recombinant proteins were purified by affinity chromatography, ion exchange chromatography and gel filtration (1L, 5L,10L, 30L, 80L,130L, 250L)≥90% purity
* SDS-PAGE

1-2 weeks

* 1~5mg recombinant protein
* Protein purification report
* Tag removal and separation of tag-free protein (if requested)

 

Service Highlights:

 

*Codon optimization: including rare codon analysis, protein hydrophilicity analysis and prediction of transmembrane domains.

*High Efficiency: our system relies on the latest technology in order to ensure high-level expression of both intracellular and secreted recombinant proteins.

*A variety of fusion tag vectors and hosts to ensure high soluble protein expression and activity.

*Scale-up: from 500ml to 500L

*Short cycle and high output

*LAL <0.1EU/mg