2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38571Ochratoxin A and patulin are mycotoxins that are produced by Aspergillus and Penicillium. They are commonly found in cereals, juice and coffee products, which cause toxic effects and cancer in human and animal. Production of antibodies and development of rapid detection methods for ochratoxin A and patulin are important issue in food safety. . Therefore, the first year objective of the present proposal is to produce the monoclonal antibodies and express ScFv (single chain variable fragment) antibody using phage display antibody system against ochratoxin A and patulin, respectively. A mycotoxin antibody library will be able to construct using this phage display antibody system. The monoclonal and ScFv will be used to establish the enzyme-linked immunosorbent assays (ELISA) and to analyze these toxins. Recently, the application of nanotechnology in biological detecting is promising. The second year objective of this proposal is focused on development of rapid immunostrip assay for both ochratoxin A and patulin. The immunostrip is using antibody-nanogold probe for detection of ochratoxin A and patulin, respectively. The assay time for immunostrip is about 5~10 min which is time saving compared with enzyme-linked immunosorbent assays. The result of immunostrip assay can be determined using naked eye without any instrument. The third year objective of this proposal is simultaneously detecting aflatoxin B1 and fumonisin B1 on a single strip based on antibody-nanogold probe. This assay will use two different antibody-nanogold conjugates and two toxins protein conjugates coating in different position on a strip membrane. Both aflatoxin B1 and fumonisin B1 can be simultaneously detected on a single immunostrip assay.赭麴毒素A(ochratoxin A)與棒麴黴素(Patulin)主要由Aspergillus與Pencillium 等常見黴菌所產生的黴菌毒素，泛存於食品與作物中，食用遭受污染的食品會導致人類許多疾病及癌症的生成。由於此類毒素對於人類及動物體健康有高度的威脅，因此開發各式抗體及建立快速免疫檢測分析法來分析調查各類食品中赭麴毒素與棒麴黴素的污染是一重要課題。因此本計劃的第一年將生產赭麴毒素A與棒麴黴素之單株抗體，並且利用噬菌體表達技術(Phage display antibody system)生產赭麴毒素A與棒麴黴素的單鏈抗體(Single chain variable fragment, ScFv)，並且建立酵素免疫分析法來檢測分析此二類毒素。此外利用此一噬菌體表達技術使用免疫鼠脾臟B細胞為抗體基因來源亦可以建立黴菌毒素抗體基因庫，篩選出針對黴菌毒素之單鏈抗體的各種專一性抗體抗體。近年來由於奈米科技的進展，利用奈米粒子於生物檢測及分析上的應用逐漸展開，因此本計畫第二年將進行赭麴毒素A與棒麴黴素奈米快速免疫檢測技術開發及應用，將奈米金粒子與抗體相結合，形成抗體奈米金探針，開發黴菌毒素的奈米快速免疫檢測試劑，以奈米金粒子與抗體相結合開發出免疫層析試紙，進行赭麴毒素A與棒麴黴素之快速檢測。本計畫第三年將利用第二年開發之技術平台，使用二種抗體在同一硝酸纖維膜膜上(nitrocellulose)膜上同時進行黃麴毒素B1與伏馬鎌孢毒素B1奈米快速檢測技術開發，將不同大小奈米金粒子分別與二種抗體相結合吸附在conjugation pad上，並且將兩種不同抗原固定硝酸纖維膜上不同位置，在同一strip上同時監測兩種黴菌毒素之新穎奈米快速免疫檢測試劑。赭麴毒素棒麴黴素噬菌體表達技術酵素免疫分析法ochratoxinpatulinphage display antibody systemELISAantibody-nanogold probeProject