CFTR Protein Domains

Accelagen Inc.

Accelagen provides comprehensive protein purification services for drug discovery and pre-clinical studies. Accelagen's protein purification expertise includes purification of full-length CFTR and many nucleotide binding domain (NBD) constructs of CFTR.

CFTR Production Services

1. Full-length recombinant CFTR

With funding and guidance from Cystic Fibrosis Foundation Therapeutics Inc. (CFFT), Accelagen has established in-house systems that enable them to express and purify full-length CFTR. With technical guidance by Dr. Jack Riordan and his colleaguesfrom the University of North Carolina at Chapel Hill, Accelagen is able to generate enough biomass essential for purification of CFTR at milligram scale, using Cell Factories to produce in large-scale stably transfected BHK cells.

2. Recombinant CFTR NBDS

Recombinant NBDs can also be made for functional and structural studies. With technical guidance by former SGX Pharmaceutical scientists, Accelagen has successfully purified the following NBD1 and NBD2 domains of CFTR:

3. Other Available Reagents

• pCDFDuet-1-GroE plasmid
• Escherichia Coli (E. Coli) BL21(DE3)GroE chemically competent cells

Certain NBD domain proteins, e.g., 5762c54BSt1pi1, are only expressed in soluble form at a significant level in the presence of chaperone co-expression, hence the above plasmid for co-expression, or competent cells harboring the chaperone protein, are generated and available to researchers.

General Services

Accelagen can provide protein expression in mammalian, baculovirus, and E. coli expression systems. Accelagen has protein purification capabilities with extensive selections of chromatography media and flexible configuration of chromatography columns tailored to each protein.

Please contact Accelagen directly regarding more detail about CFTR protein expression and purification capabilities on a fee-for-service basis.

Accelagen Inc.
6044 Cornerstone Ct W, Ste. C
San Diego, CA 92121
Tel: 858-678-8618
Fax: 888-239-6490
Toll-free: 888-365-8085

University of Alabama at Birmingham

Dr. Christie Brouillette is funded by CFFT to provide limited amounts of purified NBDs to the CF research community for pilot functional and structural studies. Typically, sub-milligram to several milligrams of pure NBDs can be provided frozen in a standard buffer containing adenosine triphosphate (ATP).

Proteins are expressed in an E. coli expression system according to the methods found in Atwell et al. (2010) PEDS 23:375-384 and Lewis et al., (2005) JBC 280:1346-1353. Plasmids for expression have been obtained from DNASU Plasmid Repository at the Arizona State University Biodesign Institute.

The following human NBD proteins are currently available. New proteins are continually added to the list so check for availability. Please contact Christie Brouillette at christie@cbse.uab.edu for additional information to request protein.

CFTR Folding Consortium

Additional protein constructs of NBD1 are available through the CFTR Folding Consortium.