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首頁> 外文會議>2010 International Conference on Optical MEMS and Nanophotonics >Development of an integrated microsystem for the multiplexed detection of protein markers in serum using electrochemical immunosensors
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Development of an integrated microsystem for the multiplexed detection of protein markers in serum using electrochemical immunosensors

機(jī)譯：使用電化學(xué)免疫傳感器對血清蛋白標(biāo)記物進(jìn)行多重檢測的集成微系統(tǒng)的開發(fā)

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Recent advances in the fabrication of microfluidic platforms initiated during the late 90s have facilitated the realisation of micro total analysis systems [1]. The integration of miniaturised fluidic handling and delivery systems with chemical and biochemical sensors provide applied scientists with powerful tools for in-field measurements away from central laboratories [2]. Amongst the various classes of elements able to transduce a chemical or biochemical events into a measurable signal, electrochemical platforms undoubtedly present the most promising advantages. Electrodes of all type, sizes and geometries can easily be integrated within a microfluidic platform and provide excellent sensitivity and versatility in comparison to other transduction techniques based on for example optical or mass sensing [3]. Furthermore, the associated electronics used to drive the electrochemical detection and signal processing can also be easily miniaturised and integrated onto the same platform by carefully designing application specific integrated circuits [4]. We have recently reported a simple and rapid approach for prototype microfluidics and sensor assembly to perform complex protein and genetic electrochemical assays with excellent reproducibility [5]. The microfluidic platform was realized by high precision milling of polycarbonate sheets, which offers flexibility and rapid turn over of the desired designs. Sixteen-electrode sensor arrays were fabricated using photolithographic deposition technologies in order to realize three-electrodes cells comprising of gold counter and working electrodes as well as silver reference electrode. Fluidic chips and electrode arrays were assembled via a laser machined double-sided adhesive gaskets, creating the microchannels necessary for sample and reagent delivery. Surface chemistry methodologies were evaluated in order to achieve the double function of eliminating non-specific binding and optimal spacing of the anchor biocomponents for m--aximum accessibility to the target proteins. Storage conditions were optimized, demonstrating a long-term stability of the reporter conjugates jointly stored within a single reservoir in the microsystem. The final system has been optimized in terms of incubation times, temperatures and simultaneous, multiplexed detection of the protein markers was achieved in less than 10 minutes with less than ng/mL detection limits. The microsystem has been validated using real patient serum samples and excellent correlation with ELISA results obtained.
機(jī)譯：在90年代末期開始的微流控平臺制造方面的最新進(jìn)展促進(jìn)了微全分析系統(tǒng)的實現(xiàn)[1]。微型流體處理和輸送系統(tǒng)與化學(xué)和生化傳感器的集成為應(yīng)用科學(xué)家提供了遠(yuǎn)離中央實驗室進(jìn)行現(xiàn)場測量的強(qiáng)大工具[2]。在能夠?qū)⒒瘜W(xué)或生物化學(xué)事件轉(zhuǎn)換為可測量信號的各種元素中，電化學(xué)平臺無疑具有最有前途的優(yōu)勢。與其他基于光學(xué)或質(zhì)量傳感的轉(zhuǎn)導(dǎo)技術(shù)相比，各種類型，尺寸和幾何形狀的電極都可以輕松集成到微流體平臺中，并具有出色的靈敏度和多功能性。此外，通過精心設(shè)計專用集成電路[4]，用于驅(qū)動電化學(xué)檢測和信號處理的相關(guān)電子設(shè)備也可以輕松地小型化并集成到同一平臺上。我們最近報道了一種用于原型微流控和傳感器組裝的簡單，快速的方法，可以執(zhí)行復(fù)雜的蛋白質(zhì)和遺傳電化學(xué)分析，并具有出色的重現(xiàn)性[5]。該微流體平臺是通過對聚碳酸酯片材進(jìn)行高精度研磨而實現(xiàn)的，它提供了所需設(shè)計的靈活性和快速的交接能力。使用光刻沉積技術(shù)制造了十六電極傳感器陣列，以實現(xiàn)由金反電極和工作電極以及銀參比電極組成的三電極電池。流體芯片和電極陣列通過激光加工的雙面膠墊圈組裝在一起，形成了樣品和試劑輸送所必需的微通道。對表面化學(xué)方法進(jìn)行了評估，以實現(xiàn)消除非特異性結(jié)合和針對m-的錨定生物組分的最佳間距的雙重功能。 -- 對靶蛋白的軸向可及性。優(yōu)化了存儲條件，證明了聯(lián)合存儲在微系統(tǒng)中單個存儲庫中的報道分子偶聯(lián)物的長期穩(wěn)定性。最終系統(tǒng)已在孵育時間，溫度和同時，不到10分鐘的時間內(nèi)以小于ng / mL的檢測限實現(xiàn)了蛋白質(zhì)標(biāo)記的多重檢測方面進(jìn)行了優(yōu)化。該微系統(tǒng)已使用真實的患者血清樣本進(jìn)行了驗證，并且與ELISA結(jié)果具有極好的相關(guān)性。

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