传感器类型
可穿戴生物传感器
检测对象
泪液葡萄糖(tear glucose, glucose);样品基质:兔眼泪液(in situ tear fluid)、体外磷酸盐缓冲液(PBS, pH 7.4)
检测原理
泪液中的葡萄糖扩散进入PMEH固定膜,被GOD识别并催化氧化,消耗氧气生成葡萄糖酸内酯和过氧化氢(H2O2)。生成的H2O2扩散至柔性Pt工作电极,在恒电位+400 mV(相对Ag/AgCl参比/对电极)下发生电化学氧化,产生与H2O2浓度成正比的电流。由于GOD反应速率受葡萄糖浓度控制,输出电流随泪液葡萄糖浓度升高而增大。PMEH(MPC/EHMA共聚物)膜用于固定GOD、减少酶泄漏并调节传质,3 wt% PMEH在灵敏度与重复性之间取得平衡。该传感器无额外化学放大,主要依靠酶催化产生H2O2和电化学换能实现连续电流读出。
检测灵敏度
校准范围: 0.03–5.0 mM;相关系数: 0.999;校准方程: 输出电流(μA)=0.245×[葡萄糖(mmol L−1)]^0.831(应用前);输出电流(μA)=0.239×[葡萄糖(mmol L−1)]^0.818(应用后)
效应效果
体外传感器响应迅速,校准范围0.03–5.0 mM,相关系数0.999,覆盖正常与糖尿病泪液葡萄糖水平。未覆膜传感器兔眼应用后性能下降;覆PMEH后重复性提高,3 wt% PMEH在灵敏度与重现性间最优,应用前后校准曲线基本不变。兔眼原位监测平均电流0.042 μA,基础泪液葡萄糖估计0.11 mM;滴入0.5 mM葡萄糖后估计0.49 mM,接近滴加浓度。口服葡萄糖耐量试验中,初始泪液葡萄糖约0.116 mM,血糖45 min达峰,泪液葡萄糖延迟约10 min、55 min达峰,最高约0.61 mM,升高约6倍,血糖升高约2倍。作者认为该软性可穿戴传感器可无创连续监测泪液葡萄糖,为血糖动态评估提供信息。
传感器的构成
- 基底/载体:PDMS软性隐形眼镜(base curve radius 8.6),提供眼表佩戴支撑与柔性。
- 柔性电极膜:70 μm PDMS薄膜,承载电极并保证弯曲不破裂。
- 工作电极:200 nm Pt薄膜,用于电化学氧化H2O2并输出电流。
- 参比/对电极:300 nm Ag与200 nm Pt双层经氯化形成Ag/AgCl,提供稳定电位与电流回路。
- 绝缘层:薄PDMS绝缘膜覆盖电极,仅暴露传感区与端子,防止短路。
- 识别元件:葡萄糖氧化酶GOD(EC 1.1.3.4),催化葡萄糖氧化生成H2O2。
- 固定/封闭膜:PMEH(MPC:EHMA=3:7共聚物)固定GOD,外覆3 wt% PMEH防止酶泄漏并提高重复性。
- 信号读出:恒电位仪施加+400 mV(vs Ag/AgCl),记录Pt电极电流。
中文摘要
本研究制备并测试了一种用于原位监测泪液葡萄糖的软性隐形眼镜生物传感器,以无创方式评估血糖。传感器采用生物相容性2-甲基丙烯酰氧乙基磷酸胆碱(MPC)聚合物和聚二甲基硅氧烷(PDMS)作为材料。通过微机电系统(MEMS)技术在PDMS膜上制备柔性铂(Pt)工作电极和氯化银/银(Ag/AgCl)参比/对电极;传感区电极修饰葡萄糖氧化酶(GOD)。体外测试中,输出电流与葡萄糖浓度在0.03–5.0 mM范围内呈良好关系,相关系数0.999,校准范围覆盖正常人和糖尿病患者报道的泪液葡萄糖浓度。将该隐形眼镜生物传感器应用于兔眼进行泪液葡萄糖监测,基础泪液葡萄糖估计为0.11 mM。通过口服葡萄糖耐量试验评估血糖变化引起的泪液葡萄糖变化,结果显示泪液葡萄糖水平较血糖水平延迟约10 min升高。结果表明,该隐形眼镜生物传感器有望提供血糖与泪液葡萄糖动态关系的更详细信息。
英文摘要
A contact lens (CL) biosensor for in situ monitoring of tear glucose was fabricated and tested. Biocompatible 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer and polydimethyl siloxane (PDMS) were employed as the biosensor material. The biosensor consists of a flexible Pt working electrode and a Ag/AgCl reference/counter electrode, which were formed by micro-electro-mechanical systems (MEMS) technique. The electrode at the sensing region was modified with glucose oxidase (GOD). The CL biosensor showed a good relationship between the output current and glucose concentration in a range of 0.03-5.0mM, with a correlation coefficient of 0.999. The calibration range covered the reported tear glucose concentrations in normal and diabetic patients. Also, the CL biosensor was applied to a rabbit for the purpose of tear glucose monitoring. The basal tear glucose was estimated to 0.11 mM. Also, the change of tear glucose induced by the change of blood sugar level was assessed by the oral glucose tolerance test. As a result, tear glucose level increased with a delay of 10 min from blood sugar level. The result showed that the CL biosensor is expected to provide further detailed information about the relationship between dynamics of blood glucose and tear glucose.