文章基本信息

标题：Optimization of an in vitro assay methodology for competitive binding of thyroidogenic xenobiotics with thyroxine on human transthyretin and albumin
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作者：Katie L. Hill ; Timo Hamers ; Jorke H. Kamstra 等
期刊名称：MethodsX
印刷版ISSN：2215-0161
电子版ISSN：2215-0161
出版年度：2017
卷号：4
页码：404-412
DOI：10.1016/j.mex.2017.10.004
语种：English
出版社：Elsevier
摘要：Graphical abstractDisplay OmittedAbstractThyroid hormones (THs) are involved in the regulation of many physiological processes in vertebrates. Competition for TH binding sites on serum transport proteins can interfere with delivery of THs to target tissues, and this is a potential mechanism of action of exogenous thyroidogenic substances. To date, detailed accounts ofin vitromethods for competitive binding with THs on TH transport proteins (human or wildlife) are sparse. In the limited number of published studies onin vitroradio-labelled TH-TH transport protein interactions, method descriptions were brief and with insufficient details for successful replication. Furthermore, upon review of these methodologies, we identified several opportunities for optimization. The present study addresses the methodological deficiencies and describes, in detail, a fully optimized and validated competitive T4 radio-ligand binding assay with human transthyretin (TTR) and albumin (ALB).•Significant improvements were made over previous methods, including better maintenance of protein stability and enhanced measurement of competition between different ligands.•Sample size was reduced to allow use of small pre-packed size exclusion chromatography columns, which eliminates the rinsing step during the separation procedure.•The assay was parameterized for use with T4 and human TTR and ALB.
关键词：Thyroid hormones;Transport proteins;Transthyretin;Albumin;Human;Competitive protein binding assay, Exogenous ligands