Expression, purification and characterization of recombinant interleukin-21

Interleukin-21 (IL-21) is a key regulator of the immune system. However, studies of this cytokine have so far been hampered by the limited availability of recombinant protein preparations. Here we describe a method based on refolding of inclusion bodies expressed in E. coli by rapid dilution. The me...

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Bibliographic Details
Published in:Journal of immunological methods 2010-10, Vol.362 (1-2), p.185-189
Main Authors: Lee, Carol M.Y., McGuire, Helen, Basten, Antony, King, Cecile, Christ, Daniel
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal - immunology
Biological and medical sciences
Cell Line
Cell Proliferation - drug effects
Chromatography, Liquid
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Gene Expression
Humans
Interleukin-21
Interleukins - biosynthesis
Interleukins - chemistry
Interleukins - immunology
Interleukins - isolation & purification
Interleukins - pharmacology
Mice
Molecular immunology
Phage display
Precursor Cells, B-Lymphoid - cytology
Precursor Cells, B-Lymphoid - immunology
Precursor Cells, B-Lymphoid - metabolism
Protein expression
Protein Folding
Protein purification
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Recombinant Proteins - immunology
Recombinant Proteins - pharmacology
Techniques
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Description
Summary:Interleukin-21 (IL-21) is a key regulator of the immune system. However, studies of this cytokine have so far been hampered by the limited availability of recombinant protein preparations. Here we describe a method based on refolding of inclusion bodies expressed in E. coli by rapid dilution. The method was applied to human and murine IL-21 proteins, which were further purified by affinity chromatography and gel-filtration. The proteins are pure and highly active as determined by endotoxin and cell proliferation assays. The availability of milligram quantities of protein enabled us to generate monoclonal antibody fragments against the cytokine and will aid in further structural, biochemical and physiological analyses.
ISSN:0022-1759
1872-7905
DOI:10.1016/j.jim.2010.08.008