學研合作計畫---功能化奈米粒子結合質譜儀運用在檢驗與診斷技術平台的開發(I)

Abstract

本計畫的總體目標為開發出一套結合功能性奈米粒子與質譜儀為基礎的檢驗與診斷 技術平台。此計畫乃基於過去六年執行國家型計畫：「功能化奈米材料於生物檢測及分 析上的應用」之研究成果而發展出來之進一步的學術研發工作，此計畫除了將進行現有 操作技術標準化及檢驗試劑工業化製備外，本計劃將擴大與成大醫院、高雄醫學大學醫 院及長庚紀念醫院研發單位合作，進行較大規模臨床樣品測試，並且與醫院現有技術進 行比較及分析，期望在未來三年，能開發出有產業化及商品化價值的產品及服務。 在過去研究成果中，我們已經成功發展出奈米探針親和性質譜方法應用於癌症標 記分子檢驗學術研究部分，這個成果主要利用奈米探針上抗體抗原交互作用和基質輔助 雷射脫附離子化時間飛行式質譜儀，可快速偵測血漿中和癌症有關的微量蛋白質，我們 已經以胃癌為技術開發目標，分析超過三百名臨床癌症病人之血液樣品，這種可多元化 設計、穩定、製程經濟、操作簡易的奈米粒子，具同時檢測多重蛋白質之優勢，預期將 可以成為一種通用的方法，成為檢測疾病標記蛋白的有效工具，取代現有ELISA 方法， 節省複雜程序，快速提供更完整的癌症臨床診斷資料。 在這個計畫的主要方向架構上，我們提出計畫建立『產業化』之具體技術平台及相 關產品： (Ⅰ) 建立功能性奈米粒子-抗體共聚物之製備技術平台及標準流程。 (II) 設計及發展商業用之免疫質譜定量檢驗之標準試劑與量測模組。 (III) 建立多重癌症標記蛋白及分析技術平台，申請相關專利。 (Ⅴ) 發展質譜分析之生物資訊軟體，建立癌症標記蛋白與臨床病人之質譜分析資料庫。 (VI) 建立癌症病人蛋白質轉譯後修飾之資料庫，開發新穎生物標記蛋白結構。 癌症早期檢測技術是現今醫學領域相當重要之議題，且具有極大之商業價值。未來 三年，將與醫院研發單位直接合作，在病人樣品及資訊取得上，有相當大之研究優勢， 且在臨床檢測認證上，醫院已有相當經驗；因此只要專利、認證及策略聯盟之研發團隊 可建立，未來計畫將申請標準檢驗認證過程 (如；FDA 檢驗認證)，以技術轉移或技術入 股方式與國內、外醫院及藥廠合作，尋求最大之『產業化』利益。

The goal of the entire project will be on the development and applications of a new molecular diagnosis platform integrating surface-functionalized nanomaterials and mass spectrometry. By the support from National Science and Technology Program for Nanoscience and Nanotechnology during the past six years, a new efficient immunoassay termed nanoprobe-based affinity mass spectrometry (NBAMS) has been established for serum protein characterization. To further implement the NBAMS for clinical nanodiagnostics, we will cooperate with Chang Gung Memorial Hospital, Kao Hsiung University Hospital and Cheng Kung University Hospital for large-scale clinical sample tests, aiming introduction of new molecular diagnosis platforms and proteomic kits in clinical diagnosis market. Based on the combination of surface functionalized nanoparticles and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, we have developed a nanoprobe-based affinity mass spectrometry immunoassay to simultaneously enrich, detect and quantify the low-abundant proteins associated with cancer. Specially, the rapid immunoassay has been tested for more than 300 patients with gastric cancer. Given the flexibility of derivation on the nanoprobes, low cost, robustness, and simplicity of the assay, our approach is advantageous in multiplexed protein detection and shows promise as superior alternative for targeted disease protein profiling. Specifically, we aim to provide the following high-content biomarker analysis service and nanoprobe kits: 1. Large-scale production and quality control of surface functionalized nanoprobe. 2. Design and development of commercialized NBAMS immunoassay kit. 3. Patent application and standard operating procedure establishment for multiplexed immunoassay in cancer diagnosis. 4. Development of informatics tool and cancer proteomics database. 5. Discovery of novel post-translational modifications (PTM) in cancer. The development of new early detection techniques for cancer diseases is of importance to human being and worthy to invest in commercial aspects. This proposed project will provide a great opportunity to bring advantage resources in Taiwan together toward the development of commercial products. In the next three years, clinical informatics, blood and tissue banks of cancer patients will be provided and tested using NBAMS technique to meet all the approval processes and to obtain the medical certifications of the local government. Also, a patent net will be filed and established based on our new technique. The technology transferring or cooperation with other research institutes and companies in the world will be executed to optimize the commercial benefit from this project eventually.