A New Method for Human Serum Phenytoin Based on Enzyme Immunoassay

概要

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A competitive binding enzyme immunoassay (EIA) for human serum phenytoin (DPH) has been developed. β-D-Galactosidase- s-_??_N-CH2CONHCH2CH2CH (C6H5) 2 prepared by reaction of N-(3, 3-diphenyl-l-propyl) maleylacetamide with β-D-galactosidase from<I>E. coli</I>was used as an enzyme labeled antigen. The insolubilized antibody used was prepared by combining yeast cell wall with immunoglobulin from sera of rabbits immunized with α, α-diphenylacetylated bovine serum albumin.<BR>After 30min competitive immunoreaction between the enzyme labeled antigen and free DPH for the insolubilized antibody, the activity of enzyme bound to the antibody was measured by using o-nitrophenyl β-D-galactoside as a substrate. One to 500ng of DPH in 10μl of sera from patients could be successfully measured by this EIA. The values of DPH concentrations measured by the EIA and the conventional gas-liquid chromatography method were found to be correlated well (r=0.96, n=100).<BR>Phenobarbital, primidone, trimethadione and carbamazepine in the sera did not interfere with the EIA, whereas 5 (4-hydroxyphenyl)-5-phenylhydantoin, when added more than 25ng per tube, interfered with the method. The method can be practically used in the ordinal clinical laboratories.
Japan Society of Clinical Chemistryの論文