分析化学2012，Vol.40Issue(4)：503-509,7.DOI:10.3724/SP.J.1096.2012.10435
核酸杂交酶联桥接分析系统定量分析寡核苷酸
Development of a Nucleic Acid Hybridization Enzyme-linked Bridging Assay System and Its Application to Oligonuleotides Quantification
孙云娟 1付洁 1王清清 1周磊 1董立厚 1欧伦 1高新 1宋海峰1
作者信息
- 1. 军事医学科学院放射与辐射医学研究所药理毒理研究室,药代动力学重点实验室,北京100850
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摘要
Abstract
An enzyme-linked bridging assay (ELBA) system based on nucleic acid hybridization principles was developed for quantification of antisense oligodeoxynucleotide (AS-ODN) in biomatrix. Conditions of the ELBA system were elaborately optimized. NeutrAvidin coated plate was used in the experiments. The optimal capture probe and detection probe concentration ratio was 2:3, Color developing time of Phosphatase Substrate (p-nitrophenyl phosphate PNPP) was 15 min and 40 U/well SI nucle-ase was used. Neither matrix nor dilution effect had influence on the quantification of ODN in plasma using ELBA, Quantitative method of the concentration of AS-ODN in monkey plasma was validated, with the quantitative linear range of 0.10-6. 25 nmol/L. The method has been successfully applied to the pharma-cokinetics(PK)profile of the AS-ODN following vein drip (v. D. )administration at 1 mg/kg,It was concluded that the hybridization-based ELBA system offered a powerful alternative tool for quantifying ODN drugs in biomatrice.关键词
寡核苷酸/药代动力学/定量/核酸杂交/酶联桥接分析系统Key words
Oligonucleotide/Pharmacokinetics/Quantification/Nucleic acid hybridization/Enzyme-linked bridging assay system引用本文复制引用
孙云娟,付洁,王清清,周磊,董立厚,欧伦,高新,宋海峰..核酸杂交酶联桥接分析系统定量分析寡核苷酸[J].分析化学,2012,40(4):503-509,7.
