Table 2 First generation (cAMP RIA assays)and second generation (cAMP/luciferase assays) for TSHR blocking autoantibodies.

	Year	Species	Assay time	Comments	Reference
FIRST generation assays (cAMP radio-immunoassay)	1978	Human	4 days	Human thyroid plasma membranes, adenyl cyclase activity	51
	1980	Human	4 days	Human thyroid plasma membranes, IgG preparation by column chromatography through DEAE-Sephadex	52
	1983	Human	4 days	Thyroid adenoma cells, IgG fractions preparation by DEAE-Sephadex column chromatography	29
	1985	Human	4 days	Human thyroid plasma membranes, IgG preparation by DEAE-cellulose column chromatography	53
	1987	Rat	4 days	FRTL-5 cells, IgG preparation by DEAE Sephadex	33
	1989	Rat	1–2 weeks	FRTL-5 cells, IgG preparation by polyethylene glycol (PEG) precipitation	68
	1989	Rat	1–2 weeks	FRTL-5 cells, IgG extraction by affinity chromatography on columns of protein A-Sepharose	66
	1990	Rat	1–2 weeks	FRTL-5 cells, IgG dialyzed in hypotonic buffer	49
	1999	Porcine	2 days	Porcine thyroid cells, IgG preparation by PEG precipitation	59
	1999	Hamster	2 days	CHO cells (clones JP02 and JP26)	73
	2000	Hamster	3 days	CHO cells (clone JP26)	55
SECOND generation assays (cAMP / luciferase)	1994	Hamster	4 days	CHO JP09 cells transfected with the recombinant human TSHR, IgG dialyzed and diluted in hypotonic buffer	17
	2001	Hamster	1 day	CHO cells (clone JP09) stably transfected with the TSHR (clone lulu*)	24
	2004, 2009	Hamster	20–24 h	CHO cells expressing the wild-type human TSHR, use of serum, spectrophotometric cAMP immunoassay	62 83
	2013	Hamster	20 h	Fresh frozen vials of CHO cells expressing the chimeric human TSHR Mc4 and a luciferase reporter gene, use of serum, luminescence (cAMP-dependent luciferase expression)	23 25 76
	2015	Hamster	8 h	CHO cells expressing the wild-type human TSHR, use of serum, no sterile conditions necessary, luminescence (cyclic nucleotide-gated calcium channel and aequorin)	63