A Chemiluminescent Immunoassay for Osteocalcin in Human Serum and a Solution to the “Hook Effect”

Table 2

Optimisation of antibody concentration.

Biotinylated antibody (μg/mL)	0.5			1			1.5
Antibody-HRP (μg/mL)	2	1	0.5	2	1	0.5	2	1	0.5
Standard 1 (0 ng/mL)	288	213	164	268	215	153	260	217	164
Standard 2 (10 ng/mL)	3009	1986	1685	2769	2145	1634	3046	2059	1748
Standard 3 (25 ng/mL)	9855	6110	5420	10151	6538	5420	10963	7126	4932
Standard 4 (50 ng/mL)	26158	17526	13341	24327	18928	13074	22867	18928	13858
Standard 5 (75 ng/mL)	35566	18850	15649	39123	18285	14710	36776	17188	14416
Standard 6 (250 ng/mL)	76489	39010	36715	80314	40961	39652	83527	44238	36083