蛋白質原位合成生物晶片之設計與製作

Design and fabrication of biochip for in-situ protein synthesis

廖仁偉(Ren-wei Liao)

指導教授 ： 楊宗勳

國立中央大學/理學院/光電科學研究所/碩士(2008年)

摘要

傳統上，蛋白質微陣列晶片其所需檢測蛋白質的量很少且種類繁多，導致前期樣品製備非常耗時且不符經濟效應。因此，本研究提出以介電質電濕式(electrowetting on dielectric, EWOD)微流體系統為基礎，利用其可驅動單一微液滴的特性，設計並製作一蛋白質原位合成生物晶片，以少量、即時和多樣化的方式合成所需的胺基酸序列，進而應用於蛋白質晶片檢測上。
本研究於使用EWOD微流體系統進行蛋白質合成的應用上，提出三點實驗結果：1. 利用加大EWOD系統中的介電層厚度以及旋轉塗布五層鐵氟龍膜，可驅動極性微流體DMF和胺基酸合成所需溶液等。2. 利用矽晶片濕蝕刻開孔的方式於鐵氟龍膜鍍上金膜，並對金膜作表面改質使其露出OH–基，使得胺基酸序列可原位接合於金膜表面。3. 由ESCA量測結果顯示，利用本研究設計之EWOD微流體系統可接合三段亮胺酸序列以及亮胺酸和苯丙胺酸兩段胺基酸序列，而每次胺基酸接合率可達五成以上。

關鍵字

胜肽合成 ； 蛋白質晶片 ； 介電質電濕式

並列摘要

Currently, the microarray chips are applied in biotechnology very popularly. All the testing samples used in the microarray are required in a small amount but in great variety, especially for the assays related to proteins. In such a way, the sample preparation will be a tedious and cost-consuming work. Therefore, how to design and to implement a biochip for in-situ synthesis of proteins efficiently becomes an urgent and attractive subject contemporarily. In this work, the electro-wetting on dielectric (EWOD) micro-fluidic system by its characteristic of driving the single drop has been applied for this work. The ESCA examination reveals the synthesis rate of the peptide is higher than 50%. It has shown that the peptides are successfully synthesized by this biochip is a small amount and the peptide sequence can be programmed in the process.

並列關鍵字

peptide synthesis ； EWOD ； protein chip

參考文獻

[1]T. Livache, H. Bazin, P. Caillat, and A. Roget, "Electroconducting polymers for the construction of DNA or peptide arrays on silicon chips," Biosensors and Bioelectronics 13, 629-634 (1998).

[2]P. J. Obeid and T. K. Christopoulos, "Continuous-flow DNA and RNA amplification chip combined with laser-induced fluorescence detection," Analytica Chimica Acta 494, 1-9 (2003).

[3]D. J. Cahill, "Protein and antibody arrays and their medical applications," Journal of Immunological Methods 250, 81-91 (2001).

[4]D. S. Mehta, C. Y. Lee, and A. Chiou, "Multipoint parallel excitation and CCD-based imaging system for high-throughput fluorescence detection of biochip micro-arrays," Optics Communications 190, 59-68 (2001).

[5]Y. Ito and M. Nogawa, "Preparation of a protein micro-array using a photo-reactive polymer for a cell-adhesion assay," Biomaterials 24, 3021-3026 (2003).