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Updated: Sep 4, 2026
A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells
Published on: July 31, 2012
[Studies on the stable integration and expression of IL-2R alpha gene in CHO cells and it's applications]
1National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.
Objectives:
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.
Methods:
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.
Results:
(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).
Conclusions:
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.
