近年來，隨著DNA晶片製作技術的成熟，蛋白質晶片在診斷及製藥方面的應用也正在研究當中。而發展蛋白質晶片的關鍵技術就在於蛋白質固化技術。然而，目前已發展的蛋白質固化技術所遭遇到的瓶頸之一為非特定性吸附。因此本研究的目的在於發展一蛋白質光固化結合自組單分子薄膜之技術，不僅能將蛋白質固定於金基材上，同時達到降低非特定性吸附的效果，並將此技術應用於蛋白質晶片之製作。本研究藉由測量接觸角、X-光電子能譜儀、原子力顯微鏡以及放射性同位素之分析來瞭解光固化流程中之表面特性。以同位素分析結果顯示，在此光固化技術中使用1 mg/ml benzophenones (BP)，蛋白質固定在金上的量為每毫米平方1.35 × 1010個分子，非特定性吸附量為28 %；若使用1 mg/ml BP將蛋白質固定在金表面上，固定量為每毫米平方1.46 × 1010個分子，並可將非特定性吸附降低至2 %以下。在應用於蛋白質晶片之製作方面，此光固化技術可藉由使用光罩曝光達到蛋白質陣列之點的大小為12 mm。另外可結合點印技術，將老鼠免疫球蛋白及33 %甘油點印在基材上與抗體反應，則可從螢光顯微鏡看到點印之大小為1 mm。從以上結果來看，此蛋白質光固化技術可成功地將蛋白質固定於金基材上，並證明此技術應用於製作蛋白質晶片之潛能及可行性。

The development of protein microarray has recently received much attention in diagnosis and pharmaceutics. A key point for the development of protein microarray is the protein immobilization technique. One of the limitations for the developed protein immobilization techniques is nonspecific adsorption. The purpose of this study was to develop the BP-based photoimmobilization combined self-assembled monolayers (SAMs) techniques for protein fixation. The SAM procedures were optimized to minimize the nonspecific adsorption and applied to the fabrication of protein microarray, including protein photopatterning via photomask and protein array with pin-printing method. Surface of SAM was characterized by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), contact angle measurement, and radiol l assay. Results show that the protein cover density was 1.46 × 1010 molecules/mm2 on the gold surface using 1 mg/ml BP-based photoimmobilibzation technique and the nonspecific adsorption could be minimized to 2 %. The patterning size of 12 mm was obtained with the illumination of UV light using photomask. Combination with pin-printing method, Mouse IgG with 33% glycerol printed to react with antibody was observed as the spot size of 1 mm. The results show that this BP based on photoimmobilization technique has the potential for the fabrication of protein array.

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