Our research is focused on

​Biological Medicine: Molecular pathogenesis of fibrotic diseases in heart, lung and kidney.
Biosensor: AuNPs biosensing platform for pathogens and biomarkers detection.
BioEnergy: Using microalgal technology to reduce CO2 and produce biomass

建立實驗小鼠模式用於檢測阻斷SARS-CoV-2/hACE2入侵肺臟細胞、導致細胞傷害之藥物開發的效能

2020年COVID-19大流行以來,造成超過四千萬人感染,超過一百二十萬人死亡。已知新型冠狀病毒(SARS-CoV-2)是透過人類第二型血管收縮素轉換酶(angiotensin converting enzyme 2, ACE2)進入肺臟細胞,進而引發炎症反應及肺組織傷害,此再度引起科學家重視ACE2的生理功能,而實驗小鼠模式的建立,可大幅加速此相關研究的進行。本實驗室早已建立ACE2 knockout (KO)小鼠,並已經由美國Jackson Laboratory引進hACE2基因轉殖小鼠(K18-hACE2 mice),因此有豐富資源進行本研究計畫。本計畫研究規劃如下:
  1. 建立mACE2 KO、hACE2 TG及hACE2 TG/mACE2 KO等三種C57BL/6J實驗小鼠,用於探討與hACE2關聯性之肺損傷及其治療策略。
  2. 建構一基因重組Lentivirus,用於表現SARS-CoV-2 spike protein S1 (a.a. 1-681),此可作為探討SARS-CoV-2的pseudovirus (Lenti-S1;可於BSL2實驗室操作)。Lenti-S1可用於感染HEK293T細胞與實驗小鼠,用於表現生產病毒S1蛋白質,並檢測此pseudovirus之in vivo感染率。
  3. 利用支氣管酸滴入方式,引起小鼠急性肺損傷之方式,建立實驗模式(acid aspiration–induced acute lung injury) (Patel et al., 2012),用於探討在Lenti-S1狀況下,ACE2與急性肺損傷修復之關聯性。
  4. 利用最新NanoBiT技術,建構一套可以在實驗動物使用的蛋白質-蛋白質交互作用的檢測系統,此用於快速、即時、精準檢測S1與hACE2接合效率。本系統可以用於檢測阻斷病毒經由hACE2感染細胞之新開發藥物的功效,包括分子藥物、soluble ACE2或S1、ACE2抗體。

利用db/db與ACE2雙基因剔除小鼠探討RAS的調節對糖尿腎病病程的影響

        Diabetes is a main cause of chronic kidney disease(CKD)that is diabetic nephropathy. Diabetic nephropathy, accounting in uremic patients with 40~50%, is one of the most important basic and clinical biomedical research topic. It is known that renin-angiotensin system (RAS) disorder induced kidney inflammation, oxidative stress, structure and fibrotic pathogenesis may relate to the disease of diabetic nephropathy through ACE-Ang II/AT1R axis and ACE2-Ang-(1-7)/Mas axis. But there are still a lot of complex molecular and pathogenic mechanisms to be clarified. Among mechanisms, it is the most worth to be explored in the new functional enzymes within RAS, such as ACE2 (angiotensin conversing enzyme II) and chymase. The function of chymase is same as ACE (angiotensin conversing enzyme), however chymase can be tissue-specifically expressed and has a higher enzyme-specific activity for the conversion of Ang I to Ang II compared with ACE. In theory, chymase-dependent Ang II-generating function and ACE2 inhibitory or antagonistic ACE(Chymase)-Ang II/AT1R axis maybe play a significant role in the pathophysiologic progression of diabetic nephropathy.
        糖尿病為慢性腎臟疾病主要病因, 此糖尿腎病(diabetic nephropathy, DN) 約占尿毒症患者之 40~50%,是當今最重要的基礎與臨床生醫研究課題之一。已知腎素 -血管收縮素系統 (renin -angiotensin system, RAS)的失調,是DN重要病程之一,此即RAS中 ACE-Ang II/AT1R axis與 ACE2-Ang-(1 -7)/Mas axis對腎臟炎症反應與纖維化病變間有密切的關聯性。據此,本研究室進行一系列 in vivo小鼠模式實驗以深入探討之,我們同時欲瞭解針對ACE、chymase及ACE2的活性調節,是否能有效地減緩DN的病程發展。 我們的研究重點有:
  1. 建立 db/db 與 ACE2 雙基因剔除小鼠模式進行DN與 RAS途徑調節的相關性研究。
  2. 探討調節腎臟組織 探討調節腎臟組織中chymase活性以控制DN病程的效應。
  3. 探討 ACE2 與 ACE2 -AngAng -(1 -7)/Mas axis在 DN 病程發展中的角色。
  4. 探討合併調節ACE/chymase和 ACE2 活性是否能更有效、且顯著減緩 DN 的病程。
本項研究的長期目標為預防或治療 DN 新藥或療法提供基礎生醫科學之依據。
 

 

PM 2.5誘發慢性阻塞肺疾病(COPD)小鼠實驗模式之建立與探討

        COPD是一種因為肺部損傷導致漸進式肺部阻塞的慢性疾病,為全球第四大死亡疾病,在未來環境中PM2.5(空氣中小於2.5微米(um)之懸浮微粒)所引起的COPD不容小覷。流行病理學研究已漸確立PM2.5對於人體健康造成的影響,其中主要為提高呼吸道疾病與死亡的風險。本研究是在此項主題研究工作包括:
  1. 汙染空氣中PM2.5的收集與分析。
  2. PM2.5處理方式與劑量對實驗小鼠的影響。
  3. 探討實驗小鼠經 PM2.5處理後呼吸系統各組織的病理病程。
我們希望藉由本實驗動物模式的建立,日後可以據此與 臨床醫師擴大合作研究的範 圍,未來還將探討不同疾病 (例如高血脂、糖、高血壓之小鼠 )對 PM2.5刺激的敏感度。
 

 

Extracellular matrix remodeling in the atrial and ventricular tissues with heart diseases

        In heart, a proper balance between synthesis and degradation of extracellular matrix (ECM) is of utmost importance in maintaining normally physiological functions. Prolonged production of several cytokines induced by wound repair process, lasting shear stress or static pressure, and challenging by reactive oxygen species can lead to excessive ECM accumulation and chronic fibrosis, often resulting in organ failure. The interplay of matrix metalloproteinases (MMPs), a family of proteolytic enzymes responsible for ECM degradation, and its naturally endogenous inhibitor TIMPs have been demonstrated to play a pivotal role in ECM remodeling processes. The pathological imbalance of MMPs and TIMPs causing abnormally accumulating ECM and fibrotic lesions has been observed in the heart failure (HF), and also in pathological atria, such as atrial fibrillation (AF). Therefore, our lab performs the studies of new insights into the underlying mechanisms involved in ECM remodeling and balance of MMPs/TIMPs in the heart diseases.
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◎ The activities of MMP-2 and MMP-9 were analyzed by gelatin-zymography
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◎ The atrial tissues of fibrillating atria (AF) and sinus rhythmus (SR) were measured by in situ zymography
 

 

​​Selected publications related to this topic

  1. Hsieh WY, Chang TH, Chang HF, Chuang WH, Lu LC, Yang CW, Lin CS*, Chang CC. 2019. Renal chymase-dependent pathway for angiotensin II formation mediated acute kidney injury in a mouse model of aristolochic acid I-induced acute nephropathy. PLoS One. 14(1):e0210656. (IF=2.776, 24/69 in MULTIDISCIPLINARY SCIENCES)
  2. Chen CY, Wu VC, Lin CJ, Lin CS, Pan CF, Chen HH, Lin YF, Huang TM, Chen L, Wu CJ; National Taiwan University Study Group on Acute Renal Failure. 2018. Improvement in Mortality and End-Stage Renal Disease in Patients With Type 2 Diabetes After Acute Kidney Injury Who Are Prescribed Dipeptidyl Peptidase-4 Inhibitors. Mayo Clin Proc. 93(12):1760-1774. (IF=7.091, 12 of 160 in MEDICINE, GENERAL & INTERNAL)
  3. Cho CC, Hsieh WY, Tsai CH, Chen CY, Chang HF, Lin CS*. 2018. In vitro and in vivo experimental studies of pm2.5 on diseases progression. International Journal Environmental Research and Public Health. 15(7). pii: E1380. doi: 10.3390/ijerph15071380. (IF=2.468, 67/185 in PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH) 
  4. Lin CI, Tsai CH, Sun YL, Hsieh WY, Lin YC, Chen CY, Lin CS*. 2018. Instillation of particulate matter 2.5 induced acute lung injury and attenuated the injury recovery in ACE2 knockout mice. International Journal of Biological Sciences 14:253-265. doi:10.7150/ijbs.23489. (IF=4.067, 82/298 in BIOCHEMISTRY & MOLECULAR BIOLOGY)
  5. Yang CW, Lu LC, Chang CC, Cho CC, Hsieh WY, Tsai CH, Lin YC, Lin CS*. 2017. Imbalanced plasma ACE and ACE2 level in the uremic patients with cardiovascular diseases and its change during a single hemodialysis session. Renal Failure 39:719-728.  (IF:1.687, 54/80 in UROLOGY & NEPHROLOGY)
  6. Hung YH, Hsieh WY, Hsieh JS, Liu FC, Tsai CH, Lu LC, Huang CY, Wu CL, Lin CS*. 2016. Alternative Roles of STAT3 and MAPK Signaling Pathways in the MMPs Activation and Progression of Lung Injury Induced by Cigarette Smoke Exposure in ACE2 Knockout Mice. International Journal of Biological Sciences 12:454-65. (IF=4.067, 82/298 in BIOCHEMISTRY & MOLECULAR BIOLOGY) Lu LC, Yang CW, Hsieh WY, Chuang WH, Lin YC, Lin CS*. 2016. Decreases in plasma MMP-2/TIMP-2 and MMP-9/TIMP-1 ratios in uremic patients during hemodialysis. Clinical and Experimental Nephrology 20:934-942. (IF=1.971, 46/80 in UROLOGY & NEPHROLOGY)
  7. Yang WJ, Liu FC, Hsieh JS, Chen CH, Hsiao SY, Lin CS*. 2015. Matrix metalloproteinase 2 level in human follicular fluid is a reliable marker of human oocyte maturation in in vitro fertilization and intracytoplasmic sperm injection cycles. Reproductive Biology and Endocrinology 13:102. (IF=2.589, 14/29 in REPRODUCTIVE BIOLOGY)
  8. Huang CH, Chang CC, Kuo CL, Huang CS, Lin CS*, Liu CS. 2015. Decrease in plasma cyclophilin a concentration at 1 month after myocardial infarction predicts better left ventricular performance and synchronicity at 6 months: A pilot study in patients with ST elevation myocardial infarction. International Journal of Biological Sciences 11: 38-47. doi: 10.7150/ijbs.10271 (IF=4.372, 72/291 in BIOCHEMISTRY & MOLECULAR BIOLOGY)
  9. Huang CH, Chang CC, Kuo CL, Huang CS, Chiu TW, Lin CS*, Liu CS. 2014. Serum iron concentration, but not hemoglobin, correlates with TIMI risk score and 6-month left ventricular performance after primary angioplasty for acute myocardial infarction. Plos ONE 9(8):e104495. (IF=3.534, 8/55 inMULTIDISCIPLINARY SCIENCES)
  10. Huang CH, Chang CC, Huang CS, Kuo CL, Chen CP, Hsia CH, Chang YM, Chen HT, Feng CC, Lin LS, Yang PT, Tsai CD, Lin CS*, Liu CS. 2013. Using oxidized low-density lipoprotein autoantibodies to predict restenosis after balloon angioplasty in patients with acute myocardial infarction. PlosONE 8(10):e74726. (IF=3.534, 8/55 in MULTIDISCIPLINARY SCIENCES)
  11. Cheng KS, Liao YC, Chen MY, Kuan TC, Hong YH, Ko L, Hsieh WY, Chen MR, Lin CS*. 2013. Circulating matrix metalloproteinase-2 and -9 enzyme activities in the children with ventricular septal defect. International Journal of Biological Sciences 9(6):557-563. (IF=4.372, 72/291 inBIOCHEMISTRY & MOLECULAR BIOLOGY)
  12. Kuan TC, Chen MY, Liao YC, Ko L, Hong YH, Yen CY, Hsieh WY, Cheng KS, Wu CL, Lin CS*. 2013. Angiotensin II downregulates ACE2-mediated enhancement of MMP-2 activity in human cardiofibroblasts. Biochemistry and Cell Biology 91:1-8. (IF=2.350, 182/291 in BIOCHEMISTRY & MOLECULAR BIOLOGY)
  13. Kuo CE, Liang SF, Lu SS, Kuan TC, Lin CS*. 2013. Estimation and prediction of drug therapy on the termination of atrial fibrillation by autoregressive model with exogenous Inputs. IEEE Transactions on Information Technology in Biomedicine 17:153-161. (IF=2.072, 22/102 in COMPUTER SCIENCE, INFORMATION SYSTEMS)
  14. Hsieh WY, Kuan TC, Cheng KS, Liao YC, Chen MY, Lin PH, Hsu YC, Huang CY, Hsu WH, Yu SY, Lin CS*. 2012. ACE/ACE2 Ratio and MMP-9 Activity as Potential Biomarkers in Tuberculous Pleural Effusions. International Journal of Biological Sciences 8:1197-1205.
  15. Kuan TC, Yang TH, Wen CH, Chen MY, Lee IL, Lin CS*. 2011. Identifying the regulatory element for human angiotensin-converting enzyme 2 (ACE2) expression in human cardiofibroblasts. Peptides 32;1832-1839.
  16. Chang CC, Kuan TC, Hsieh YY, Ho YJ, Sun YL, Lin CS*. 2011. Effects of diosgenin on myometrial matrix metalloproteinase-2 and -9 activity and expression in ovariectomized rats. International Journal of Biological Sciences 7:837-847.
  17. Lin CS*, Pan CH, Wen CH, Yang TH, Kuan TC. 2010. Regulation of angiotensin converting enzyme II by angiotensin peptides in humancardiofibroblasts. Peptides 31:1334-1340.
  18. Lin LM, Lai LP, Lin CS, Chou NK, Chiu CY, Lin JL. Left ventricular extracellular matrix remodeling in dogs with right ventricular apical pacing.Journal of Cardiovascular Electrophysiology 2010:1142-1149.
  19. Chen CL, Huang SK, Lin JL, Lai LP, Lai SC, Liu CW, Chen WC, Wen CH, Lin CS*. 2008. Upregulation of matrix metalloproteinase-9 and tissue inhibitors of metalloproteinases in rapid atrial pacing-induced atrial fibrillation. Journal of Molecular and Cellular Cardiology 45:742-753.
  20. Lin CS*, Pan CH. 2008. Regulatory mechanisms of atrial fibrotic remodeling in atrial fibrillation. Cellular and Molecular Life Sciences 65:1489-1508.
  21. Pan CH, Wen CH, Lin CS*. 2008. Interplay of angiotensin II and angiotensin 1-7 in the regulations of matrix metalloproteinases of humancardiocytes. Experimental Physiology 93:599-612.
  22. Lin CS, Lai LP, Lin JL, Sun YL, Hsu CW, Chen CL, Mao SJT, Huang SKS. 2007. Increased expression of extracellular matrix proteins in rapid atrial pacing-induced atrial fibrillation. Heart Rhythm 4:938-949.
  23. Pan CH, Lin JL, Lai LP, Chen CL, Huang SKS, Lin CS*. 2007. Downregulation of angiotensin converting enzyme II is associated with pacing-induced sustained atrial fibrillation. FEBS Letters 581:526-534.
  24. Chen CL, Lin JL, Lai LP, Pan CH, Huang SKS, Lin CS*. 2007. Altered expression of FHL1, CARP, TSC-22 and P311 provide insights into complex transcriptional regulation in pacing-induced atrial fibrillation. Biochimica et Biophysica Acta 1772:317-329.

提升微藻養殖技術與價值,開發與產製微藻生物活性物

        ​Photosynthetic organisms include plants, algae and some photosynthetic bacteria. These organisms use solar energy to combine water with CO2 to create biomass. Moreover, high oil-producing plant and algae can be used to produce biodiesel. Algae are more efficient at using CO2 for photosynthesis, have a higher proliferation rate, and can be more readily incorporated into engineered systems. Furthermore, microalgaltechnology provides a means for recycling waste carbon from fossil fuel combustion. Algal biodiesel is one of the only avenues available for high-volume re-use of CO2 generated in power plants. Therefore, in our planning, the focus of research is the production of biodiesel from high lipid-content algae utilizing waste CO2 from the burning of fossil fuels. The major missions include:
  1. collection of oil-producing algae.
  2. construction of photobioreactor for biomass production of microalgae.
  3. advances in our understanding on the physiology and biochemistry of algae for biomass- and oil-production.
  4. establishment of close or open systems for recycling of waste CO2.
  5. oil extract and purification of cultured algae.
  6. biological conversion of algal oil to biodiesel.
 

 
微藻 (microalgae)具高效能光合作用,可有效率利用陽光,將CO2轉換成生物能 (biomass),微藻並富含許多生物活性質,是近年來深具研發價值之主題。本研究室持續與成功大學、中山大學、農業科學 院、金屬研究中心、中國鋼鐵公司、台糖公司合作致力於微藻減碳、生質能源、水資源再利用等研究 ,而長期 研究目標包括:
  1. 建構大規模單元操作的Photobioreactor/Raceway Raceway模組(10 與 20 噸規模),研發提升微藻養殖效能之操作模式。
  2. 篩選特種微藻株, 其除具有高效率生長特性外, 並富含特定生物活性物。
  3. 誘發微藻生產特定生物活性物質之養殖模式,探討其抑菌、抗氧化、 細胞修復、提升免疫力等之生物功能性。
  4. 開發能用於飼料添加能用於飼料添加 、產製機能性健康食品之製品 。
 

 
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Selected publications related to this topic

  1. Kuo CM, Jian JF, Sun YL, Lin TH, Yang YC, Zhang WX, Chang HF, Lai JT, Chang JS, Lin CS*. 2018. An efficient PBRs/Raceway circulating system combined with alkaline-CO2 capturing medium for microalgal cultivation. Bioresource Technology 266: 398-406. (IF=6.669, 13/162 in BIOTECHNOLOGY & APPLIED MICROBIOLOGY)
  2. Lee TM, Tseng YF, Cheng CL, Chen YC, Lin CS, Su HY, Chow TJ, Chen CY, Chang JS. 2017. Characterization of a heat-tolerant Chlorella sp. GD mutant with enhanced photosynthetic CO2 fixation efficiency and its implication as lactic acid fermentation feedstock. Biotechnology for Biofuels 10:214. (IF=5.452, 18/162 in BIOTECHNOLOGY & APPLIED MICROBIOLOGY)
  3. Kuo CM, Lin TH, Yang YC, Zhang WX, Lai JT, Wu HT, Chang JS, Lin CS*. 2017. Ability of an alkali-tolerant mutant strain of the microalga Chlorella sp. AT1 to capture carbon dioxide for increasing carbon dioxide utilization efficiency. Bioresource Technology 244:243-251. (IF=6.669, 13/162 in BIOTECHNOLOGY & APPLIED MICROBIOLOGY)
  4. Ho SH, Chiu SY, Kao CY, Chen TY, Chang YB, Chang JS, Lin CS. 2017. Ferrofluid-assisted rapid and directional harvesting of marine microalgal Chlorella sp. used for biodiesel production. Bioresource Technology 244:1337-1340. (IF=6.669, 13/162 in BIOTECHNOLOGY & APPLIED MICROBIOLOGY)
  5. Kuo CM, Jian JF, Lin TH, Chang YB, Wan XH, Lai JT, Chang JS, Lin CS*. 2016. Simultaneous microalgal biomass production and CO2 fixation by cultivating Chlorella sp. GD with aquaculture wastewater and boiler flue gas. Bioresource Technology 221:241-250. (IF=6.669, 13/162 in BIOTECHNOLOGY & APPLIED MICROBIOLOGY)
  6. Kuo CM, Chen TY, Lin TH, Kao CY, Lai JT, Chang JS, Lin CS*. 2015. Cultivation of Chlorella sp. GD using piggery wastewater for biomass and lipid production. Bioresource Technology 194:326-333. (IF=6.669, 13/162 in BIOTECHNOLOGY & APPLIED MICROBIOLOGY)
  7. Chiu SY, Kao CY, Chen TY, Chang YB, Kuo CM, Lin CS*. Cultivation of microalgal Chlorella for biomass and lipid production using wastewater as nutrient resource. Bioresource Technology (In press; doi:10.1016/j.biortech.2014.11.080) (IF=5.039, 1/12 in AGRICULTURAL ENGINEERING)
  8. Kao CY, Chen TY, Chang YB, Chiu TW, Lin HY, Chen CD, Chang JS, Lin CS*. 2014. Utilization of carbon dioxide in industrial flue gases for the cultivation of microalga Chlorella sp. Bioresource Technology 166:485-493. (IF=5.039, 1/12 in AGRICULTURAL ENGINEERING)
  9. Kao CY, Chiu SY, Huang TT, Dai1 L, Ling-Kang Hsu, Lin CS*. 2012. Ability of a mutant strain of the microalga Chlorella sp. to capture carbon dioxide for biogas upgrading. Applied Energy 93:176-183. (IF=5.261, 6/133 in ENGINEERING, CHEMICAL)
  10. Kao CY, Chiu SY, Huang TT, Dai1 L, Wang GH, Tseng CP, Chen CH, Lin CS*. 2012. A mutant strain of microalga Chlorella sp. for the carbon dioxide capture from biogas. Biomass & Bioenergy 36:132-140. (IF=3.411, 2/12 in AGRICULTURAL ENGINEERING)
  11. Chiu SY, Kao CY, Huang TT, Lin CJ, Ong SC, Chen CD, Chang JS, Lin CS*. 2011. Microalgal biomass production and on-site bioremediation of carbon dioxide, nitrogen oxide and sulfur dioxide from flue gas using Chlorella sp. culture. Bioresource Technology 102:9135-9142. (IF=5.039, 1/12 inAGRICULTURAL ENGINEERING)
  12. Ong SC, Kao CY, Chiu SY, Tsai MT, Lin CS*. 2010. Characterization of the thermal-tolerant mutants of Chlorella sp. with high growth rate and application in outdoor photobioreactor cultivation. Accepted by Bioresource Technology. Bioresource Technology 101:2880-2883.
  13. Chiu SY, Tsai MT, Kao CY, Ong SC, Lin CS*. 2009. The air-lift photobioreactors with flow patterning for a high-density culture of microalgae and carbon dioxide removal. Engineering in Life Science 9:254-260.
  14. Chiu SY, Kao CY, Tsai MT, Ong CS, Chen CH and Lin CS*. 2009. Lipid accumulation and CO2 utilization of Nannochloropsis oculata in response to CO2 aeration. Bioresource Technology 100:833-838.
  15. Chiu SY, Kuan TC, Kao CY, Ong SC, Chen CH, Lin CS*. 2008. Reduction of CO2 by a high-density culture of Chlorella sp. in asemicontinuous photobioreactor. Bioresource Technology 99:3389-3396.

Patent
  1. Lin CS, Chiu SY, Kao CY. Microalgae cultivation module. IPA-ROC I433647 (專利權期間:2014/4/11~ 2031/9/5)
  2. Lin CS, Kao CY. Microalgae cultivation module including circular tube-shaped cultivation bag. IPA-ROC: I457073 (利權期間:2014/10/21~ 2032/7/18)
  3. Cheng JS, Lin CS, Chen CY, Kao CY. Carbon dioxide-capturing apparatus for microalgae cultivation and the method thereof. IPA-ROC: I479988 (專利權期間:2015/4/11~ 2032/8/27)
  4. Tseng CP, Chen CC, Lin CS. Biogas electric generator and electricity generation method using microalgae carbon capture. (Pending IPA-ROC: 101107655)
  5. Lin CS, Kao CY, Chen TY, Chang YB, Chiu SY. The technique of magnetic particles applied to microalgae harvest. (Pending IPA-ROC: 102149050)
利用奈米金球建構食品微生物與蛋白質酶活性之生物感測平台
近年來,生物感測技術逐漸被應用於醫學、食品、農業、環境等多方面領域。而本研究室也積極發展生物感測技術平台,其中利用奈米金球(gold nanoparticles; AuNPs)於生物分子檢測,以大幅提昇生物感測技術的敏感性與可應用性為我們研究的核心。本研究室陸續發展質量式生物感測器-石英晶體微天平(quartz crystal microbalance; QCM)、網版印刷碳電極試片(screen-printed carbon electrode; SPCE)及AuNPs效應等生物分子檢測平台,其皆具備操作簡便,檢測時間短,檢測樣本需求量低,以及高靈敏度等優勢。本研究室在此領域的研究成果主要刊登於Biosensors and Bioelectronics、Nanoscale 以及BiomaterialsNanotechnologyFood Microbiology、Journal of Agricultural and Food Chemistry 等期刊。相關的研究成果已分別獲得2個中華民國、1個美國及1個大陸發明專利。
 

 

A gold nanoparticles optical biosensing platform used to assay proteinase activity

        The surface plasmon resonance (SPR) wavelength of colloidal gold nanoparticles (AuNPs) would vary when the AuNPs aggregate, possess different sizes, shape, or being modified with chemical molecules.  In this study, an optical biosensing platform for the assay of proteinase activity was established based on the SPR property of AuNPs. The 13-nm AuNPs were modified with gelatin (AuNPs-gelatin) as proteinase substrate and then modified with 6-mercaptohexan-1-ol (MCH) (AuNPs/MCH-gelatin). After proteinase (trypsin or gelatinase) digestion, the AuNPs lost shelter and MCHincreases the attractive force between modified AuNPs. Therefore, the AuNPs close to each other gradually and resulting in AuNPs aggregation. TheAuNPs aggregation could be monitored by a red-shift of surface plasmon absorption and a visible color change of the AuNPs was from red to blue. Such color change could be observed with naked eyes. In the detection, absorption ratio of A625 nm/A525nm of the reacted AuNPs solution could be used to quantitatively estimate the proteinase activity, and a linear correlation has been established when the trypsin activity was from 1.25 × 10-1 to 1.25 × 102U and the matrix metalloproteinase-2 activity was from 50 ng/mL – 600 ng/mL.
 

 

Electrochemical biosensing system

Fabricating the Screen-printed carbon electrodes (SPCE) biosensor for pathogens detection:
        The field of electrochemical biosensor has grown rapidly in recent years. Electrochemical sensors of detecting virus infection and bacterial contamination have also been developed. In our lab, a disposable amperometric immunosensor strip was fabricated for the rapid detection of Dengue virus and E. coli O157:H7. Screen printed carbon electrodes (SPCE) were framed by commercial silver and carbon ink, and then electrochemically characterized by coupling them with HRP-conjugated antibody. Hydrogen peroxide and ferrocenedicarboxylic acid (FeDC) were used as the HRP substrate and mediator, respectively. Response current (RC) of the immunosensor strips could be significantly amplified by 13-nm Au nanoparticles (AuNPs) immobilized on the working electrode. Our results show that the combined effects of AuNPs and FeDC enhanced RC approximately 13 folds. Our studies point out that the SPCE modified with AuNPs has potential as further applications to incorporate into an integrated system for rapid pathogen detection.
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Selected publications and patents related to this topic

  1. Quintela IA, de los Reyes BG, Lin CS, Wu VCH. 2019. Simultaneous colorimetric detection of a variety of Salmonella spp. in food and environmental samples by optical biosensing using oligonucleotide-gold nanoparticles. Frontiers in Microbiology 10:1138. doi: 10.3389/fmicb.2019.01138 (IF=4.259, 32/133 in Microbiology)
  2. Chang HF, Sun YL, Yeh FY, Tseng IH, Chang CC, Lin CS*. 2018. Detection of chymase activity using the specific peptide probe conjugated onto gold nanoparticles. RSC Advances 8:29013 – 29021. (IF=3.049,69/172 in CHEMISTRY, MULTIDISCIPLINARY)
  3. Sun YL, Wu HT, Lin CS. 2016. Direct colorimetric detection of canine distemper virus by RT-PCR combined with gold nanoparticles. IOSR Journal of Biotechnology and Biochemistry 2(6):69-74.
  4. ​Quintela IA, de los Reyes BG, Lin CS, Wu VCH. 2015. Simultaneous Direct Detection of Various Shiga-toxin Producing Escherichia coli (STEC) Strains by optical biosensing with oligonucleotide-functionalized gold nanoparticles.  Nanoscale 7:2417-2426. (IF=7.394, 21/259 in MATERIALS SCIENCE, MULTIDISCIPLINARY)
  5. Yeh FY, Liu TY, Tseng IH, Yang CW, Lu LC, Lin CS*. 2014. Gold nanoparticles conjugates-amplified aptamer immunosensing screen-printed carbon electrode strips for thrombin detection. Biosensors and Bioelectronics 61:336-343. (IF=6.451, 1/27 in ELECTROCHEMISTRY)
  6. Yeh FY, Tseng IH, Chuang SH, Lin CS*. 2014. Spacer-enhanced chymotrypsin-activated peptide-functionalized gold nanoparticle probes: a rapid assay for the diagnosis of pancreatitis. RSC Advances 4:22266-22276. (IF=3.708, 38/148 in CHEMISTRY, MULTIDISCIPLINARY)
  7. Chuang YC, Huang WT, Chiang PH, Tang MC, Lin CS*. 2012. Aqueous zymography screening of matrix metalloproteinase activity and inhibition based on colorimetric gold nanoparticles. Biosensors and Bioelectronics 32:24-31. (IF=6.451, 1/27 in ELECTROCHEMISTRY)
  8. Guo X, Lin CS (contribution as the first author), Chen SH, Ye R, Wu VCH. 2012. A piezoelectic immunosensor for specific capture and enrichment of viable pathogens by quartz crystal microbalance sensor, followed by detection with antibody-functionalized gold nanoparticles.Biosensors and Bioelectronics 38:177-183. (IF=6.451, 1/27 in ELECTROCHEMISTRY)
  9. Chu PT, Lin CS, Chen WJ, Chen CF, Wen HW. 2012. Detection of gliadin in food using a quartz crystal microbalance biosensor that incorporates gold nanoparticles. Journal of Agricultural and Food Chemistry 60:6483-6492. (IF=3.107, 2/56 in AGRICULTURE, MULTIDISCIPLINARY)
  10. Chuang YC, Li JC, Chen SH, Liu YU, Kuo CH, Huang WT, Lin CS*. 2010. An optical biosensing platform for proteinase activity using gold nanoparticles. Biomaterials 31:6087-6095. (IF=8.312, 2/77 in ENGINEERING, BIOMEDICAL)
  11. Chen SH, Chuang YC, Lu YC, Lin HC, Yang YL, Lin CS*. 2009. A method of layer-by-layer gold nanoparticles hybridization in a quartz crystal microbalance DNA sensing system used to detect dengue virus. Nanotechnology 20:215501.
  12. Chen SH, Lin KI, Tang CY, Peng SL, Chuang YC, Lin YR, Wang JP, Lin CS*. 2009. Optical detection of human papillomavirus type 16 and type 18 by the sequence sandwich hybridization with oligonucleotide-functionalized Au nanoparticles. IEEE Transactions on NanoBioscience8:120-131.
  13. Lin YH, Chen SH, Chuang YC, Lu YC, Shen TY, Chang CA, Lin CS*. 2008. Disposable amperometric immunosensing strips fabricated by Au nanoparticles-modified screen-printed carbon electrodes for the detection of foodborne pathogen Escherichia coli O157:H7. Biosensors and Bioelectronics 23:1832-1837.
  14. Chen SH, Wu VCH, Chuang YC, Lin CS*. 2008. Using oligonucleotide-functionalized Au nanoparticles to rapidly detect foodborne pathogens on a piezoelectric biosensor. Journal of Microbiological Methods 73:7-17.
  15. Wu VCH, Chen SH, Lin CS*. 2007. Real-time detection of Escherichia coli O157:H7 sequences using a circulating-flow system of quartz crystal microbalance. Biosensors and Bioelectronics 22:2967-2975.

Patent
  1. Chen SH, Lin CS, Chen GT, Lin YH, Shen TY. Composite modified electrode trip. IPA-ROC: I336782 (專利權期間:2011/2/1~2027/7/4; 交大與五鼎生技公司共擁專利所有權)
  2. Chen SH, Lin CS, Chen GT, Lin YH, Shen TY. Composite modified electrode trip. US 8236154 (專利權期間:2012/8/7~2027/7/4; 交大與五鼎生技公司共擁專利所有權)
  3. Chen SH, Lin CS, Chen GT, Lin YH, Shen TY. Composite modified electrode trip. CN: 101339154 (專利權期間:2013/3/27~2027/7/4; 交大與五鼎生技公司共擁專利所有權)
  4. Lin CS, Yen FY, Tseng IF. Nanoparticle fluorescent probe for detecting protease activity and use thereof. IPA-ROC:I500769 (專利權期間:2015/9/21~ 2033/12/29)