Production of Recombinant Chemokines and Validation of Refolding

Abstract

The diverse roles of chemokines in normal immune function and many human diseases have motivated numerous investigations into the structure and function of this family of proteins. Recombinant chemokines are often used to study how chemokines coordinate the trafficking of immune cells in various biological contexts. A reliable source of biologically active protein is vital for any in vitro or in vivo functional analysis. In this chapter, we describe a general method for the production of recombinant chemokines and robust techniques for efficient refolding that ensure consistently high biological activity. Considerations for initiating development of protocols consistent with Current Good Manufacturing Practices (cGMPs) to produce biologically active chemokines suitable for use in clinical trials are also discussed.

Keywords: Chemokine; Chemokine oxidation; Chemokine purification; Chemokine refolding; Expression; Mass spectrometry; Mass spectrometry of chemokines; NMR; Protein; Protein NMR; Protein folding; Purification; Recombinant chemokines; Recombinant cytokines; Refolding; cGMP.