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Eukaryotic expression and biological activity of human interleukin-35 / 中国医学科学院学报
Acta Academiae Medicinae Sinicae ; (6): 618-622, 2013.
Artigo em Chinês | WPRIM | ID: wpr-285950
ABSTRACT
<p><b>OBJECTIVE</b>To express and purify recombinant human interleukin-35[IL-35-IgG1 (Fc) in eukaryotic expression system and to study the interaction of IL-35 with gp130 protein.</p><p><b>METHODS</b>A mammalian expression vector, pSTEP2-IL35-LFc, was constructed and transfected into HEK293T cells. Then rhIL-35-IgG1 (Fc) was expressed and purified with protein A affinity chromatography, and was examined with SDS-PAGE and Western blot analysis. The binding of IL-35 to its receptor gp130 was investigated using enzyme-linked immunosorbent assay (ELISA). The biological effect of IL-35 on gp130 was explored in M1 myeloid leukemia cells.</p><p><b>RESULTS</b>rhIL-35-Fc with high purity on reduced SDS-PAGE was obtained. ELISA confirmed that IL-35-Fc was bound to gp130 and neutralized the function of gp130 in M1 myeloid leukemic cells.</p><p><b>CONCLUSIONS</b>High-purity and biologically active rhIL-35-Fc protein successfully produced in this study. IL-35 binds to gp130 and neutralizes its activity of in M1 myeloid leukemic cells.</p>
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Ensaio de Imunoadsorção Enzimática / Transfecção / Western Blotting / Interleucinas / Metabolismo Limite: Humanos Idioma: Chinês Revista: Acta Academiae Medicinae Sinicae Ano de publicação: 2013 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Ensaio de Imunoadsorção Enzimática / Transfecção / Western Blotting / Interleucinas / Metabolismo Limite: Humanos Idioma: Chinês Revista: Acta Academiae Medicinae Sinicae Ano de publicação: 2013 Tipo de documento: Artigo