設施農業：智慧感測【生機系吳靖宙特聘教授】
論文篇名 英文：Electrochemical sandwich immunoassay for quantification of therapeutic drugs based on the use of magnetic nanoparticles and silica nanoparticles.
中文：使用奈米磁珠與奈米矽珠以電化學三明治免疫分析法對治療藥物的定量檢測
期刊名稱 Journal of Electroanalytical Chemistry
發表年份,卷數,起迄頁數 2019, 849, 113381
作者 Ming-Jie Lin, Yi-Ming Chen, Chen-zhong Li, Ching-Chou Wu*
DOI 10.1016/j.jelechem.2019.113381
中文摘要 Adalimumab(Ada)抗體藥物已被廣泛用於自身免疫疾病的治療。正確的Ada劑量可以維持治療效果與減輕症狀。本研究發展了以奈米磁珠(MNPs)免疫反應與可拋式網印碳電極()檢測技術用以快速定量Aa的方法。抗Ada的Fab片段與老鼠的抗人抗體分別被使用為一級抗體(Ab1)與二級抗體(Ab2)。葡萄糖氧化酵素(GOD)與Ab2以濃度3:1比率被共同固定於奈米矽珠(SiNPs)，以維持足夠的免疫親和力與較大的GOD數量。當Ada依序與Ab1@MNP、GOD-Ab2@SiNPs免疫反應後，所形成的GOD-Ab2@SiNPs/Ada/Ab1@MNPs以磁場分離，並回溶後滴到塗布glucose/ferricyanide的SPCE上以GOD進行催化，被還原成ferrocyanide的氧化電流可用以定量Ada。該MNP-based的免疫感測器在0.1-2 μg/mL濃度有良好的線性範圍，其檢測極限為15.5 ng/mL。此免疫分析法可實際量測10倍稀釋之病人血清中的Ada濃度，具有良好的準確率，整體的操作時間約0.5 h，此拋棄式MNP-based免疫分析技術未來可用於治療性藥物監控的量化平台。
英文摘要 Adalimumab (Ada) is widely used to treat autoimmune diseases, and a correct dose of Ada can maintain therapeutic effectiveness and relieve symptoms. This study develops a rapid quantifying method of Ada using magnetic nanoparticle (MNP)-based immunoreaction and disposable screen-printed carbon electrodes (SPCEs). Anti-Ada Fab fragment and mouse anti-human antibody is respectively used as primary antibody (Ab1) and secondary antibody (Ab2). Glucose oxidase (GOD) and the Ab2 were co-immobilized on silica nanoparticles (SiNPs)with the concentration ratio of 3:1 to promise good immune affinity and the more amount of immobilized GOD. After sequentially immunoreacting Ada with Ab1@MNPs and GOD-Ab2@SiNPs, the separated GOD-Ab2@SiNPs/Ada/Ab1@MNPs complex was dripped on the glucose/ferricyanide-coated SPCEs for GOD catalysis, and then the anodic current of reduced ferrocyanide was used for the quantification of Ada. The MNP-based immunoassays have good linearity from 0.1μg/mL to 2μg/mL and a limit of detection of 15.5 ng/mL. Moreover, the immunoassays can practically measure the Ada concentrations in 10-fold diluted patient serum samples with good recovery. The operation time of MNP-based immunoassays takes only about 0.5 h. The disposable MNP-based immunoassays have great promise to develop a rapid quantification platform for therapeutic drug monitoring.