- My protein needs very specific co-factors or buffer conditions!
Special requests can be communicated when you submit your target online. However, we recommend that you first talk to a member of the PPP to verify the feasibility of your request before you submit your target for production.
- I donít think E. coli will be able to produce my protein!
There is evidence that as many as 80% of human soluble targets have at least one domain that can be expressed in E. coli. While we do realize that this might not help you, we still believe that the multi-construct approach in E. coli is the right place to start your trials. However, we have certainly noted lower success rates for proteins larger than 80 kD, membrane associated proteins or proteins with complex PTMs. If your protein falls into any of these categories, we could give it a try in insect and/or mammalian cells. Talk to us!
- Where are you located?
We are located at NTU School of Biological Sciences, 60 Nanyang Drive (we were previously at NTU@one-north hosted by IMCB/A-STAR in the Proteos building).
In-silico to in-vitro
Use the LIMS to create and manage your protein constructs
The platform will clone your protein constructs into E. coli
High-throughput protein purification
Your protein constructs will be purified using IMAC and SEC
Purification using affinity chromatography
All constructs will be fused to a N-terminal cleavable HIS6-tag
Expression in insect cells for difficult proteins
We are currently implementing high-throughput cloning using baculovirus
Transmembrane proteins will be screened for optimal detergent conditions
Multiple quality control steps
All protein batches will be verified using mass spectrometry and/or sequencing
Receive plasmids, expression strain glycerol stocks and protein batches