Small scale protein expression screening and optimization
Reliable, reproducible and correctly folded soluble recombinant protein production method is necessary for most structural biology, functional biology, drug discovery and enzymology projects. Protein expression and proper folding depends on several factors 1. Vector used 2. E.coli strain 3. Expression conditions such as temperature, inducing agent, composition of media (carbon source ), induction time etc 4. Fusion tags - Histag, GST tag, MBP tag etc 5. Membrane protein. Whether you need few micrograms or several milligrams of protein we can help to optimize the expression in E.coli. We can screen several conditions in parallel and optimize the expression conditions for the existing construct without the need for subcloning into a different vector. Once the right expression condition is identified we also offer to scale up and do process development and large scale production from 5 liters to 400 liters.
Deliverables:
- Comparison of the SDS page gel image for the different conditions tested - total cell lysate, both the soluble and pellet fractions are run against molecular weight markers and compared with control sample.
- Best condition identified for soluble protein expression.
Pricing:
- Our prices start from $500 and up and depends on the number of expression conditions, E.coli strains and the need for any subcloning.
To help you optimise expression levels, improve protein solubility and protein stability, and decrease protein toxicity we have put together 4 different packages that has already been tested and found successful by our scientists. You can also choose 1 or more packages or a combination of packages.
Package A - Expression of your protein under 4 different temperature conditions - 15°C, 20°C, 30°C and 37°C.
Package B -Expression of your protein which is already cloned in a modified pET vector in 4 different commercially available E.coli strains- BL21(DE3), BL21 (DE3) pLysS, BL21 (DE3) CodonPlus-RIL (-RP) or Rosetta (DE3) or Origami (DE3).
Package C -Protein expression using different carbon sources - LB media, 2x YT, Superbroth and autoinduction using glucose as the sole source of carbon.
Package D - Subcloning into different vectors - with a selection of fusion tags such as GST, MBP, Thioredoxin or Nus A and checking protein expression after addition of fusion tags.
If you have someother expression condition in mind, please contact us with your needs and we will certainly help you in designing your project. This email address is being protected from spambots. You need JavaScript enabled to view it.