[Studies on the stable integration and expression of IL-2R alpha gene in CHO cells and it's applications].
OBJECTIVE
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.
Cheng
X
X
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.
Du
H
H
Zhang
Y
Y
Gao
J
J
Heng
F
F
Wu
N
N
Shen
Y
Y
D000818:Animals; D016466:CHO Cells / Q000378:metabolism; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D015870:Gene Expression; D006801:Humans; D007376:Interleukin-2 / Q000032:analysis*; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D007376:Interleukin-2; D015375:Receptors, Interleukin-2