Cheng X, Du H, Zhang Y, Gao J, Heng F, Wu N, Shen Y
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2000 Apr;22(2):130-3.
To establish a novel bioassay method for quantitative analysis of human IL-2 based on the specific binding of Interleukin-2 receptor alpha subunit (IL-2R alpha) with IL-2.
Southern blot hybridization was first used to detect the stability of integration of recombinant secretive IL-2R alpha (rsIL-2R alpha) gene into the genome of highly expressed cell line reported elsewhere; the apparent Mr of the rsIL-2R alpha was then determined by using Western blotting; finally, a receptor-antibody sandwich ELISA method has been established for quantitative analysis of IL-2.
(1) Stable integration of rsIL-2R alpha gene into the genome of #17 CHO cell line has been identified; (2) the apparent Mr of rsIL-2R alpha was approximately 40,000; (3) linear range of the standard curve obtained from the receptor-based ELISA fell between 31.25-500 U of IL-2 (r = 0.9995). The slop of the standard curve decreased significantly when IL-2 was pre-incubated with goat anti-IL-2 antibody IgG (P < 0.01).
An IL-2R alpha-based IL-2 ELISA has been established for laboratory and clinical use with advantages of accuracy, specificity and simplicity over other conventional bioassays for IL-2 detection.
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