Large-scale Protein Preparation Large-scale Protein Preparation

Large-scale production of recombinant proteins is necessary to produce the quantities of protein for detailed analyses, i.e., probing for protein-ligand interactions, producing antibodies, characterizing enzymatic properties, and facilitating structural studies by X-ray crystallography or NMR spectroscopy. This requires expressing the protein at the optimized conditions, which should yield tens of milligrams of target protein per liter of culture. The Core is equipped with an 11-L fermenter to provide the culture volume and with chromatography equipment to purify such amounts of proteins. We have also conducted large-scale refolding of proteins from inclusion bodies.

Below is an example of a protein produced by large-scale induction and its purity after affinity chromatography and concentration. Presented with kind permission from Dr. E. Spicer, Biochemistry and Molecular Biology, MUSC.

 

CD Spectrum indicating the protein is structured

 

SDS-PAGE at different stages during production and purification

1. unpurified sample
2. affinity purified sample
3. marker
4. final concentrated Sample

 

 

 

 

 

 

Contacts Contacts

Director: Yi-Te Hsu, Ph.D. Associate Professor
hsuy@musc.edu
Tel: 843-792-0849
Co-Director: Erika E. Büllesbach, Ph.D. Associate Professor
bullesee@musc.edu Tel: 843-792-9926
Research Specialist II: Mr. Dzmitry Fedarovich
fedarov@musc.edu Tel: 843-792-6529