Information & Contact
The Protein Expression Facility (PEF) is a multi-user facility which serves primarily faculty and students from the laboratories in the Kasha Laboratory Building. In addition, laboratories in Biology, Biochemistry, Chemistry and Nutrition perform projects in the Facility. To learn more, contact PEF Director, Dr. Gwi Moon Seo.
Types of projects
The primary purpose of the PEF is to facilitate the large scale expression of recombinant proteins in eucaryotic cells. We have methods and equipment for that purpose, including the WAVE bioreactor and the BelloCell bioreactor. Other projects include expression of native proteins in response to chemical or biochemical inducers or metabolic pressures, uptake of nanoparticles and dyes, production of siRNA knockout gene clones and analysis of effects, and cytological studies of the cellular location of novel proteins.
Special techniques and capabilities
Expertise includes growth of virus with host cells, both insect (Baculovirus) and mammalian (SV40, adenovirus, AAV), isolation and purification of the viruses, immunoassay development, transient transfection of mammalian cells, and selection and isolation of stable cell clones; growth assays for measurement of stimulatory effects of factors; cell analysis (viable cell density, cell size, cell shape) using a CedexHiRes. Most techniques involving cultured eucaryotic cells can be handled in the facility.
Initiating a project in the PEF
The process begins with an initial meeting with PEF director involving the principal investigator and the individual who will perform the work in the facility. A plan will be laid out for the scope and extent of the proposed work and an order form for the work will be filed. This will allow the facility to have adequate materials on hand for the project and provide documentation for the end of the month billing. A training schedule for the worker will be negotiated at the same time.
Working in the Facility
The lab is a biosafety level 2 (BSL2) laboratory designated by FSU Environmental Health and Safety, according to the CDC/NIH Joint Guidelines on Biosafety in the Biomedical Laboratory. The suite has been designed specifically to facilitate cell culture projects by multiple users- in both culture rooms which are isolated from all others and in common areas, such as the microscopy area and analytical area where users can gather to compare and discuss results. In addition to the entire suite being isolated, each culture room is also isolated containing all necessary equipment and supplies for culture within the room. Four biosafety cabinets continuously filter the air; six incubators allow isolation of cultures by bio-safety level. A manual of biosafety rules and regulations guides behavior and introduces users to biosafety equipment and design.
Protein Expression Facility Fee Structure
In November 2009 all of the IMB/KLB Core Facilities will begin assessing fees to partially defray the cost of the Facilities. The fees for PEF represent only the actual cost of materials. Please consult the PEF User Fees schedule to determine the user contribution for various types of work performed at the Facility.
| Service | Price |
|---|---|
| Bacterial Expression | |
| E.coli expression trial, per construct | $80.50 |
| Protein expression and production from E.coli 2L culture | $117.50 |
| Protein expression and production from E.coli, additional 2L (up to 24L) | $12.00 |
| Cell pellet production, E.coli, per liter | $34.70 |
| Baculovirus Expression | |
| Insect cell pellet production, per liter | $169.50 |
| Baculovirus production per construct/50 ml | $5.25 |
| Protein production in insect cells from 1L | $206.50 |
| Mammalian expression | |
| Expression trial, per construct | $90.75 |
| Protein production in mammalian cell from 1L | $216.75 |
| Suspension culture protein expression, additional liter | $48.50 |
| Protein Purification | |
| Single step | $74.50 |
| Two steps | $127.00 |
| Three steps | $186.50 |
| Endotoxin removal | $100.00 |
| Tag cleavage with protease | $45.50 |
| Additional services | |
| Cell counter (Countess 2) | $2.50 |
| Competent cell (BL21, BL21(DE3),DH5alpha, Top10, Rosetta(DE3), SoluBL21, XL10-gold, Nico21(DE3), BL21-Codonplus(DE3)-RIPL, DH10B, DH10Bac | $0.65 |
| Proteases (Ulp1, 3C protease) | $2.63 |
| Enzymes (T7 RNA Polymerase) | $2.63 |
* For a cost estimate for your external project, please contact the facility director.