文章基本信息

标题：A One-Step Enzyme-Linked Immunosorbent Assay for a Novel Osteoblast Differentiation-Promoting Compound, TAK-778 in Serum
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作者：Tomofumi KUROKAWA ; Tetsuo HOSHINO ; Tsuneo ODA 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：1999
卷号：22
期号：12
页码：1266-1270
DOI：10.1248/bpb.22.1266
出版社：The Pharmaceutical Society of Japan
摘要：TAK-778, ((2R, 4S)-(-)-N-[(4-diethoxyphosphorylmethyl)phenyl]-1, 2, 4, 5-tetrahydro-4-methyl-7, 8-methylene-dioxy-5-oxo-3-benzothiepin-2-carboxamide), is a novel osteoblast differentiation-promoting compound, and its sustained-release formulation is expected to be clinically used for the enhancement of fracture healing. To date, TAK-778 levels in serum have been measured using conventional reverse-phase HPLC with inferior sensitivity and time-consuming procedures. We have produced polyclonal antibodies against TAK-778 by using one of its derivatives coupled with a carrier protein, and developed a one-step ELISA for the determination of TAK-778 in serum. The antibodies had minimal cross-reactivities to the biologically inactive metabolites including the oxidized-form (0.36%) and the cleavaged-form (0.00083%). The competitive ELISA was accomplished within three hours with a detection limit of 0.5ng/ml serum, and the coefficients of variations for samples ranging from 1 ng/ml to 80 ng/ml were 1.1-4.4% in an intra-assay and 3.1-9.6% in an inter-assay. The present ELISA is so rapid, sensitive and selective for TAK-778 that it could be conveniently used in a clinical field for the determination of many serum specimens.
关键词：ELISA;TAK-778;osteoblast differentiation;antibodies