心血管疾病（CVD）是世界上死亡率和發病率最高的疾病。現今的心血管疾病(例如:心肌梗塞（MI）和心力衰竭（HF）)臨床檢測是利用對血液中蛋白質生物標記濃度進行評估。然而現今的檢測技術，需要昂貴且複雜的實驗室大型儀器來進行，檢測過程耗時且成本高昂。針對此問題，我們擬開發一快速診斷工具，將半導體技術和生物醫學工程結合，開發一生物感測器晶片。此生醫感測晶片技術將提供快速且便宜的心血管疾病篩檢方式。在此項研究中，我們致力開發的生物感測晶片具備便攜性、高敏感度、無須繁雜的操作步驟及免除試劑的優點。能在5分鐘內檢測人類血液中的蛋白質生物標記濃度。此晶片技術採用電雙層（EDL）場效電晶體（FET）生物感測器，此設計克服尋常FET感測器於高鹽度環境中電荷屏蔽的限制，在人體鹽類濃度環境下仍具備良好的感測靈敏度。
此研究中研發了兩種類型生物感測晶片：FET嵌入式感測晶片及和柵極延伸FET感測器晶片。FET嵌入式感測晶片將測試溶液與FET通道接觸影響電訊號。 柵極延伸FET感測器晶片則將感測區域建於延伸柵極的金屬電極，將感測區域上的免疫化學反應轉化為電訊號輸出。更高的電場輔助選通晶體管的漏極電流可以提高FET感測晶片於全血檢測中的靈敏度和選擇性，並使FET感測晶片具備更低的檢測極限和更寬的濃度檢測範圍。此感測晶片的感測過程，通過在將一滴血滴在感測晶片上，接著把裝置向下翻轉，血液中的細胞將因重力與血漿分離。在5分鐘的感測時間後，便攜式傳感器系統測量感測器輸出電信號，並將測量結果呈現在個人電腦中。我們已經在標準生理緩衝液中成功檢測人血清與全血中的心肌肌鈣蛋白I（cTnI），利尿鈉肽（BNP和NT-proBNP）和D-二聚體等心血管疾病相關生物標記。此技術透過市場調查以及商業評估，認為此技術具備低成本及操作簡單的優點。我們的心血管疾病生醫感測晶片檢測能由任意人員操作並在5分鐘內獲得檢測結果。適合在家庭，辦公室和醫院等場所運用。此外，FET生物感測器平台用於生物組織的電學表徵，證明我們的感測器平台在外科手術輔助工具中的潛在應用。
為了進一步研究感測器表面的物理現象以及更好的感測器結構設計。我們針對不同感測器結構設計影響進行了調查，並將最佳傳感的關鍵設計參數確定為電極間距離，傳感區域，表面功能化和測試溶液介電強度。使用交流電阻抗分析技術建立感測器模型調查信號轉導中的電容性質。
總結此項研究:一、一種新型EDL門控FET生物感測器，其性能優於傳統的FET生物感測器。二、使用EDL門控FET生物傳感器進行全血蛋白質生物標記診斷。三、使用EDL門控FET生物感測器的生物組織表徵工具。四、建立EDL門控FET生物感測器的信號轉導機制。五、研究感測器相關機制，以增強使用EDL門控FET生物感測器對蛋白質檢測的靈敏度。

In this age of scientific and technological revolution, the diseases that can be easily managed/cured are still proving fatal, with increased mortality and morbidity rates, which do not seem to improve. Cardiovascular diseases (CVDs) are the foremost reasons of mortality and/or morbidity in the world. Medical intervention carried out for CVDs such as myocardial infarction (MI) and heart failure (HF) are based on the assessment of the amount of certain molecular biomarkers like proteins from blood, which necessitate complex and laboratory bound instruments that are expensive, labor intensive and time-consuming. For developing a rapid diagnostic or screening tool, we focused on integrating semiconductor technology and biomedical engineering, on a single biosensor chip that would facilitate consumer based biomedical applications. Through this research, we primarily aim to develop a portable and sensitive biosensing platform to detect protein biomarkers present in whole blood, in the span of 5 minutes, by eliminating the use of any extensive sampling steps or additional reagents. The sensing approach employs an electrical double layer (EDL) gated field effect transistor (FET) biosensor, which has improved sensitivity in high ionic strength test media, overcoming the fundamental limitation of charge screening in traditional FET biosensors. Two types of sensing structures are investigated: FET embedded sensor structure, where the test solution comes in contact with FET channel during signal transduction; and extended gate structure, in which a pair of gold based electrodes intended as extended gate assemblies, translate the immunochemical reactions at the sensor surface into electrical output. Higher electric field assisted gating of the drain current of transistor allows improved sensitivity and selectivity, low detection limit and a wide dynamic range of detection, in whole blood. By placing just a drop of blood on the device, the cellular components found in blood are gravitationally separated from the blood plasma during the 5 minutes of incubation time period, by inverting the device’s orientation downwards. A portable sensor system is devised to measure the output signals from the sensor and present the measurement results in personal computer integrated with the measurement unit. We have demonstrated the detection of clinically relevant cardiac markers such as cardiac troponin I (cTnI), natriuretic peptides (BNP and NT-proBNP) and D-dimer in standard physiological buffer, human serum and whole blood. The sensor technology has been established to significantly bring down the overall cost and complexity of the assay, with nominal user intervened protocols. The whole blood assay developed using our sensor can be executed at the accessibility of homes, offices and hospitals, by persons with or without prior training, in just 5 minutes. Furthermore, the FET biosensor platform is utilized for electrical characterization of biological tissues, to elucidate potential applications of our sensor platform in surgical assistance tools. To further elucidate the physical phenomenon at the sensor interface, an investigation into the sensor response at the wake of different structural design considerations is carried out and critical design parameters for optimal sensing are identified as inter-electrode distance, sensing area, surface functionalization and test solution dielectric strength. AC impedance analysis is used to develop a sensor model to encompass the capacitive nature of signal transduction. To summarize, this research describes: i. a novel EDL gated FET biosensor that outperforms the traditional FET biosensors, ii. a whole blood protein diagnostic assay using EDL gated FET biosensor, iii. a biological tissue characterization tool using EDL gated FET biosensor, iv. modeling of signal transduction mechanism of EDL gated FET biosensor and v. design guidelines to enhance sensitivity of protein detection using EDL gated FET biosensor.

Chapter 1. Introduction 9
1.1 Motivation 9
1.2 Project Goal 12
Chapter 2. Literature Review 17
2.1 Molecular Biomarkers 17
2.2 Point of care and home-care diagnostics 19
2.3 Field effect transistor (FET) based biosensor 22
2.3.1 AlGaN/GaN high electron mobility transistor (HEMT) 33
Chapter 3. Experimental Design 37
3.1 AlGaN/GaN HEMT fabrication 37
3.2 Fabrication of extended gate sensor array chip 40
3.3 Surface functionalization 41
3.4 Measurement of sensor 44
3.5 Protein elution process 44
Chapter 4. Results and Discussion 46
Section 4.1. Electrical Double Layer (EDL) gated FET Biosensor: Structure and mechanism 47
4.1.1 Sensor design 47
4.1.2 Sensor working 49
Section 4.2. Protein biomarker detection using contact mode HEMT embedded biosensor 57
4.2.1 Objective 57
4.2.2 Contact mode EDL gated HEMT embedded biosensor for Cardiac troponin I detection 61
4.2.2.a Antibody based cTnI detection 61
4.2.2.b Aptamer based cTnI detection 69
4.2.2.c Portable biosensor system for high sensitivity cTnI assay 72
4.2.3 Contact mode EDL gated HEMT embedded biosensor for BNP detection 77
4.2.3.a Detection of purified BNP in 1X PBS with 4% BSA 77
4.2.3.b Detection of BNP in human serum 78
Section 4.3. Whole Blood Diagnostics using Extended gate EDL FET Biosensor 81
4.3.1 Objective 81
4.3.2 Background response of whole blood on extended gate EDL FET biosensor 82
4.3.3 cTnI detection using extended gate EDL FET biosensor 92
4.3.3.a Detection of purified cTnI in 150 mM standard buffer solution with 4% BSA 92
4.3.3.b Detection of cTnI spiked in whole blood 95
4.3.3.c Detection of cTnI in clinical whole blood 98
4.3.4 BNP detection using extended gate EDL FET biosensor 102
4.3.4.a Detection of purified BNP in 1X PBS with 4% BSA 102
4.3.4.b Detection of BNP spiked in whole blood 103
4.3.4.c Detection of BNP in clinical whole blood 104
4.3.5 NT-proBNP detection using extended gate EDL FET biosensor 107
4.3.5.a NT-proBNP detection in 1X PBS with 4% BSA 108
4.3.5.b Sensor characteristics in whole blood 111
4.3.5.c Sensitivity as a function of surface receptor immobilization 116
Section 4.4. Sensor mechanism 123
4.4.1 Objective 123
4.4.2 Effect of electric field strength on sensitivity 124
4.4.3 Design considerations of the liquid gated FET sensor 127
4.4.3.a Investigation of liquid gated FET drain current characteristics 127
4.4.3.b AC impedance analysis for investigation of sensor design 134
4.4.4 Protein detection in different sensor design 145
4.4.5 Sensor model 149
4.4.6 Protein detection in different ionic strength 157
Section 4.5. Electrical characterization of biological tissue 161
4.5.1 Objective 161
4.5.2 Single pulse measurement of biological tissue 163
4.5.3 Biphasic pulse measurement of biological tissue 168
Chapter 5. Future Work 174
Chapter 6. Summary 179
Reference 181

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