Development of a sandwich enzyme-linked immunosorbent assay (ELISA) to quantify γ-glutamyl-carboxylated clotting factor IX and assess redox susceptibility of anticoagulant chemicals

开发夹心酶联免疫吸附试验 (ELISA) 来量化 γ-谷氨酰羧化凝血因子 IX 并评估抗凝化学物质的氧化还原敏感性

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作者:Ryo Sato, Kanami Watanabe, Ryo Kamata, Kazuki Takeda

Abstract

Anticoagulant chemicals (ACCs) such as warfarin are widely used in medical applications as well as for their rodenticide properties. Their efficacy is greatly influenced by polymorphisms in the gene encoding vitamin K epoxide reductase (VKOR). Evaluation of the activity of ACCs toward VKOR variants is essential to determine their proper use. Presently, this is achieved by co-expressing VKOR of Rattus Norvegicus and human clotting factor IX in cultured cells and measuring inhibition of vitamin K-dependent gamma-glutamyl carboxylation of factor IX (glaFIX) activity. However, glaFIX has only been quantified using indirect methods like blood coagulation assays. We have developed a sandwich enzyme-linked immunosorbent assay using a glaFIX-specific antibody to quantify glaFIX and used this to analyze inhibition of VKOR activity by warfarin.

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