BioMedical Engineering Laboratory

Gold Nanoparticels Biosencing System

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Biomaterials、Nanotechnology、Food 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)