Services
Protein expression and purification
Taking an expression system constructed in the Investigator’s lab, the lab will develop a protocol to produce the protein on a large scale (typically, at the milligram level). The investigator should provide all available information about the recombinant protein, including details of the expression system used, the sequence of the protein and any known properties, e.g. whether it is membrane-bound, post-translational modifications etc. The method used to assay the protein (e.g. enzyme assay, Western etc) should also be described.
The Protein Science Lab will perform a large-scale expression and purify the protein using one or more chromatographic steps (e.g. Ni 2+-affinity, ion exchange or gel filtration). The investigator lab will receive the purified product together with a written report (including details of the protocol used).
Charges are levied accordingly to the complexity of the purification procedure, i.e. the number of chromatographic steps required to obtain the desired purity or whether refolding is required etc.
For new proteins, a one-time charge is made to conduct a trial expression and purification, and to develop a protocol for purification.
To date, the lab has used only E. coli expression systems and is not yet properly equipped for yeast, baculovirus or mammalian cell expression. As the lab expands, however, these services will be added.
Other services
Cloning: The lab functions best when the expression system has already been developed in the investigator’s lab, but, depending on the current work load of the lab, requests to clone the gene of interest into expression vectors (for a negotiated fee) will be considered. If time does not allow, then the lab will advise on the best strategies for cloning and can provide a suitable expression vectors for use by the investigator.
Circular dichroism: The lab can also determine the folded state of the protein using circular dichroism (for an additional fee).
Crystallization: If a crystal structure of the protein of interest is desired, the lab can also set up a number of crystallization trials (fee levied).