Molecular biology. The lab has a wide collection of commercial and lab-customized expression vectors for bacterial and yeast strains. It can provide services such as primer design, gene cloning by RT-PCR, sequence verification, site-directed mutagenesis, expression vector development based on customer request, subcloning the target gene into various expression vectors and protein expression in different host cells or cell-free systems.

Computer-controlled fermentation. The lab has two New Brunswick Bioflo 3000 bioreactors that can be operated from 1-L to 10-L scale and a 150-L Sartorius BBI bioreactor. Fermentation services include equipment preparation and sterilization, batch or fed-batch fermentation, isotopic labeling of expressed proteins and cell harvesting to yield a concentrated cell paste. The lab has developed several robust control strategies for different fermentation requirements. Variables that may be controlled to optimize protein yield include media composition, pH, nutrient composition and supplement feed rate, temperature, inducer concentration, dissolved oxygen concentration, induction time, and harvest time.

Downstream processing. The lab provides post-fermentation processing services, including cell concentration using a Millipore Pellicon tangential flow filtration system or a Sorvall centrifuge, cell lysis using an Avestin Emulsiflex C3 high pressure homogenizer or sonication, and freeze-drying using a Labconco Lyph Lock 4.5 unit. Protein purification is performed using a GE Healthcare ÄKTA Basic UPC 100 high performance liquid chromatography system. The protein purity and expression level can be evaluated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), total-protein assay, western blotting and/or activity assays (e.g., enzymatic assays). A walk-in cold room is available for processing temperature-sensitive proteins. Cell mass, crude protein or purified product can be freeze dried for preservation.

Additional fermentation services. Unused fermentation capacity in the PEL could be made available to make other products. Advanced process control schemes may be implemented to optimize product formation. The availability of fermentation capacity at the 1 L, 10 L, and 150 L scales provides a fermentation scale-up capability rarely found in academia.

 

For more information or work order, please contact the lab director or lab manager.