Production of Recombinant Chemokines and Validation of Refolding

Christopher T. Veldkamp, Chad A. Koplinski, Davin R. Jensen, Francis C. Peterson, Kaitlin M. Smits, Brittney L. Smith, Scott K. Johnson, Christina Lettieri, Wallace G. Buchholz, Joyce C. Solheim, Brian F. Volkman

Research output: Chapter in Book/Report/Conference proceedingChapter

13 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages539-565
Number of pages27
DOIs
StatePublished - 2016

Publication series

NameMethods in Enzymology
Volume570
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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  • Cite this

    Veldkamp, C. T., Koplinski, C. A., Jensen, D. R., Peterson, F. C., Smits, K. M., Smith, B. L., Johnson, S. K., Lettieri, C., Buchholz, W. G., Solheim, J. C., & Volkman, B. F. (2016). Production of Recombinant Chemokines and Validation of Refolding. In Methods in Enzymology (pp. 539-565). (Methods in Enzymology; Vol. 570). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2015.09.031