Detection of thyroid-stimulating antibody using frozen stocks of Chinese hamster ovary cells transfected with cloned human thyrotropin receptor.
To measure thyroid-stimulating antibody (TSAb) in sera from patients with Graves' disease, we developed a new assay system with using frozen stocks of CHO-K1 cells. CHO-K1 cells transfected with cloned thyrotropin (TSH) receptor on a 96 well plate were frozen in Cell Banker and stored at -70 degrees C. Three days before the assay, they were thawed in the culture medium and allowed to grow in a monolayer until use. The medium was replaced with medium containing IgGs from the patients, then after 2 h, it was collected and concentrations of adenosine 3'-5'-cyclicmonophosphate (cAMP) were measured. This method is sensitive enough to detect TSAb and it is simpler and easier than the methods which use FRTL-5 cells.
Takano
T
T
Department of Laboratory Medicine, Osaka University Medical School, Japan.
Sumizaki
H
H
Amino
N
N
D000818:Animals; D016466:CHO Cells / Q000378:metabolism*; D002460:Cell Line; D006224:Cricetinae; D015925:Cryopreservation; D003470:Culture Media; D006801:Humans; D018828:Immunoglobulins, Thyroid-Stimulating / Q000032:analysis*; D051381:Rats; D011989:Receptors, Thyrotropin / Q000235:genetics* / Q000276:immunology; D012680:Sensitivity and Specificity; D014162:Transfection
D003470:Culture Media; D018828:Immunoglobulins, Thyroid-Stimulating; D011989:Receptors, Thyrotropin