利用表面電漿子共振矽基片B型抗諧振反射光波導生物感測元件於抗原抗體鍵結之檢測

Detection of the Binding between Antibody and Antigen by ARROW-B SPR Biosensors

王家駿(Chia-Chun Wang)

指導教授 ： 袁俊傑 ； 崔秉鉞

國立交通大學/資訊學院/生醫工程研究所/碩士(2015年)

https://doi.org/10.6842/NCTU.2015.00589

摘要

本論文研究以B型抗諧振反射光波導(ARROW-B)表面電漿子共振(SPR)生物感測器量測抗體抗原之間的鍵結。有別於以往利用螢光標定於抗原抗體之偵測,本生物感測器有免標定、即時偵測、低成本與步驟簡單快速之優點。抗體的檢測為醫學上常見的疾病篩檢的方式,但由於傳統檢測方式耗時過長,故希望利用本元件提供疾病篩檢上一個更好選擇。本元件利用表面電漿 子共振現象作為生物感測的方式,當感測區域中液體等效折射率的改變時,會造成通過波導的光強度訊號有所變化。而在波導表面所使用的金金屬,與生物分子可以透過巰基十一酸(11-MUA)作為連結,其巰基與金表面會形成硫 金強鍵結。當巰基十一酸自組裝分子層穩定後,依序通入抗原與抗體,從即時偵測的結果可以觀察到每個階段皆有明顯的響應變化。實驗結果顯示,該 感測器可以明確的量測到抗原抗體之間的鍵結,進而驗證生物學中已知現象來確認元件之可行性。

關鍵字

表面電漿子共振 ； 型抗諧振反射光波導 ； 生物感測器 ； 抗體

並列摘要

In this study, we detected the binding between antibody and antigen by using antiresonant reflecting optical waveguide B (ARROW-B) surface plasmon resonance (SPR) biosensors. Compared with traditional fluorescence methods, our sensor has many advantages including label-free, real-time, low cost and rapid process. This sensor was designed for the aqueous environment, and the SPR phenomenon is the core principle we utilized for detection. Detection of antibodies is a common way to check up diseases, but traditional detections spend more time than our method. Therefore, we hope to offer a better choice to people with ARROW-B SPR sensors. When we injected different reagents into the chamber, it caused the changes of effective index, giving rise to variations of light power. An Au-coated surface can be linked with biomolecular by 11-Mercaptoundecanoic acid (11-MUA), besides the bond between gold and 11-MUA is strong. After self-assembled monolayer of 11-MUA had formed, we sequentially injected antigen and antibody. Each step caused obviously changes in power detection. The experimental results show that the binding of antibody and antigen can be detected by our sensor. Moreover, we verified known phenomenon in biology to affirm the feasibility of ARROW-B SPR sensors.

並列關鍵字

surface plasmon resonance ； antiresonant reflecting optical waveguides ； ARROW-B ； SPR ； biosensor ； antibody

參考文獻

[1] Borghesi L, Milcarek C, "From B cell to plasma cell: regulation of V(D)J recombination and antibody secretion," Immunol. Res, vol. 36, pp. 27--32, 2006.

[2] Parker D, "T cell-dependent B cell activation," Annu Rev Immunol, vol. 11, pp. 331--360, 1993.

[3] Stern P, "Current possibilities of turbidimetry and nephelometry," Klin Biochem Metab, vol. 14, pp. 146--151, 2006.

[4] J. Hunter, Treatise on the Blood, Inflammation and Gunshot wounds. Pennsylvania: Webster, 1794.

[6] S. Cohen and R. R. Porter, "Structure and biological activity of immunoglobulins," Adv. Immunol, vol. 3, pp. 287--349, 1964.