電阻式C反應蛋白感測器之研發

指叉式生物感測器具有快速診斷、省時、敏感性高的優點，成為目前生物感測器的研發主軸，透過阻抗式電訊號的直接量測，希望能取代傳統的分子檢驗技術或是臨床生化檢驗。CRP不僅是急性發炎時的標記，也是心血管疾病的指標。本研究以網印的方式製備出銀碳複合電極，利用不同的苯環衍生物( Hydrocinnamic acid、2-Phenylethanethiol、Phenethylamine)做為媒介，透過交聯劑(ethyl(dimethylaminopropyl) carbodiimide /N-Hydroxysuccinimide、Glutaraldehyde )以共價鍵結方式將anti-CRP固定在電極表面，藉此增加固定化程序的穩定度。在掃描式電子顯微鏡觀察下，電極表面結構變得較為粗糙，表示電極表面有被化學藥劑修飾；元素含量結果中分析出硫、氮或氧表示抗體是有成功固定在電極表面上；蛋白質定量上可知電極接枝率大約在80%左右；而循環伏安法測量結果顯示，經過苯環衍生物修飾後之電極有促進電子傳遞與放大感測訊號的功能，可提升感測器之靈敏度；電化學阻抗譜的結果顯示，當接上CRP蛋白之後會阻礙電極表面電子的傳遞，導致阻抗變大的情形發生，此證明電極有很好的鍵結能力，能夠固定蛋白質。本研究製備出之網印電極，經由mediator修飾，可以將anti-CRP穩定的固定在電極表面上，增加的電化學反應和固定分子之數量，因此非常有潛力發展成生物感測器。

關鍵字

C-反應蛋白；循環伏安法；阻抗分析儀

並列摘要

Impedance spectroscopy has become the main research topic in the development of biosensor due to the advantages of rapid diagnosis, faster detection time and high sensitivity. Through direct measurement of impedance telecommunications allows the replacement of traditional biochemical tests. C-reactive protein is an inflammatory marker believed to be of value in the prediction of coronary events. The purpose of this study is to develop a highly selective and specific impedance biosensor for CRP detection. A silver-carbon electrode was prepared by screen printing method, namely screen-printed electrode (SPE). The electrode was then immersed in benzene-derived compounds such as hydrocinnamic acid, 2-phenylethanethiol or phenethylamine solution, followed by immobilization of anti-CRP antibody on the surface of electrode via covalent bonding with cross-linking agen. SEM observation showed that the surface of SPE became rough after treated with anti-CRP, indicating that the electrode had been successfully modified. Element content analysis showed that sulphur, nitrogen and oxygen was specifically recognized on the surface of SPE. The SPE has grafting rate of about 80% when connected with anti-CRP antibody. The measurement of cyclic voltammetry (CV) showed that benzene-derived modified SPE would have enhanced sensor sensitivity. Electrochemical impedance spectroscopy (EIS) showed that protein CRP would hindered the electron transfer on the surface of SPE and lead to higher impedance, which proved that the electrode has high binding affinity of CRP. Using benzene-derived compounds as the mediator could increase the electroactive area and the amount of immobilized biomolecules. The SPE system we designed has the potential to develop into biosensors.

並列關鍵字

C-Reactive Protein ； cyclic voltammetry ； impedance spectroscopy

參考文獻

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國際替代計量

電阻式C反應蛋白感測器之研發

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