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.
Article
in Chinese
| 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>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Enzyme-Linked Immunosorbent Assay
/
Transfection
/
Blotting, Western
/
Interleukins
/
Metabolism
Limits:
Humans
Language:
Chinese
Journal:
Acta Academiae Medicinae Sinicae
Year:
2013
Type:
Article
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