The PECF provides protein refolding services. The following strategy has been implemented to identify conditions under which insoluble proteins may be refolded into their native conformations. The investigator supplies an E. coli cell pellet from an induced culture (1L) along with SDS-PAGE results that confirm high level expression. The PECF will:
Isolate inclusion bodies from cell paste.
Perform primary refolding screening to identify starting parameters.
Perform additive screening to optimize conditions. Extra additives (protein dependent) can be tested at this time if desired.
Scale up refolding under optimized conditions with 5-10 mg of inclusion body protein to provide material for purification and subsequent structural and/or biological studies.
The presence of correctly folded protein can be evaluated by CD spectroscopy within the PECF.
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