title: Efficient production of recombinant IL-21 proteins for pre-clinical studies by a two-step dilution refolding method.
abstract: Produced by CD4(+) helper T cells and natural killer T (NKT) cells, interleukin-21 (IL-21) performs broad regulatory functions on B cells, CD4(+) T cells, CD8(+) T cells, NK cells and NKT cells. Targeting IL-21 to enhance the immune system has attracted great interests in the development of vaccination, anti-infection and anti-tumor therapies. Administration of IL-21 in pre-clinical models is however limited by relatively high expense of the recombinant IL-21 protein. Here, we report a rapid and cost-effective method to produce IL-21 using Escherichia coli (E. coli) by introducing a novel two-step dilution strategy for refolding. The method has been validated to produce milligrams of human IL-21, human IL-21/IL-4 chimera and mouse IL-21 with high bioactivities and low endotoxin, mostly suitable for in vitro and in vivo pre-clinical studies.
authors.affiliation: Department of Immunology (Clayton), School of Biomedical Sciences, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Zhian
authors.initials: Z
authors.lastname: Chen
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Cui
authors.forename: Yew Ann
authors.initials: YA
authors.lastname: Leong
authors.forename: Travis
authors.initials: T
authors.lastname: Beddoe
authors.forename: Di
authors.initials: D
authors.lastname: Yu
mesh_terms: D000818:Animals; D002460:Cell Line; D004353:Drug Evaluation, Preclinical; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D053738:Interleukin-21 Receptor alpha Subunit / Q000378:metabolism; D007378:Interleukins / Q000096:biosynthesis* / Q000737:chemistry; D051379:Mice; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D000098546:Interleukin-21
chemical_list: C508117:IL21R protein, human; C508118:Il21r protein, mouse; D053738:Interleukin-21 Receptor alpha Subunit; D007378:Interleukins; D011994:Recombinant Proteins; D000098546:Interleukin-21