title: Soluble expression of recombinant human CD137 ligand in Escherichia coli by co-expression of chaperones.
abstract: CD137 ligand (CD137L) is a member of the tumor-necrosis factor superfamily that binds CD137 to provide positive co-stimulatory signals for T cells activation. Co-stimulation through CD137/CD137L has become one of the promising approaches for cancer therapy. Previous reports have shown that CD137L expressed in Escherichia coli resulted in inclusion bodies or low yield. In this study, the effects of five different chaperone teams on the soluble expression of recombinant human CD137L protein were explored and analyzed. The poor expression of CD137L in the cytoplasm of E. coli was improved significantly by co-expression of chaperone GroES-GroEL-Tf. After dual induction and affinity chromatography, purified recombinant CD137L was obtained at a yield of 3 mg protein per liter with purity greater than 98% from original undetectable level. Additionally, the purified recombinant CD137L could bind CD137-positive cells in a dose-dependent manner, markedly promote the growth of activated mice T cells, and elevate the release of IL-2. The present work provides an effective system for soluble expression of functional human co-stimulatory molecule CD137L, which will facilitate the clinical developments of recombinant protein drugs.
authors: Laboratory of Chemical Biology, China Pharmaceutical University, Nanjing, China.
authors: Shuzhen
authors: S
authors: Wang
authors: Aimin
authors: A
authors: Tan
authors: Junfang
authors: J
authors: Lv
authors: Peng
authors: P
authors: Wang
authors: Xiaojin
authors: X
authors: Yin
authors: Yijun
authors: Y
authors: Chen
mesh_terms: D053330:4-1BB Ligand / Q000378:metabolism*; D000818:Animals; D057005:Bioengineering; D001709:Biotechnology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D006801:Humans; D008213:Lymphocyte Activation / Q000502:physiology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018832:Molecular Chaperones / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D053330:4-1BB Ligand; D018832:Molecular Chaperones; D011994:Recombinant Proteins