单株抗体-国立宜兰大学生物技术与动物科学系.pptVIP

4.7 Preserving Proteins Lyophilization (freeze-drying) Protein, usually a liquid product, is first frozen A vacuum is used to hasten the evaporation of water from the fluid Will maintain protein structure and can be stored at room temperature for long periods of time 4.8 Scale-Up of Protein Purification Protocols are usually designed in the laboratory on a small scale Must be scaled up for production Process is approved by FDA so must make sure laboratory procedures can be scaled up 4.9 Postpurification Analysis Methods Protein Sequencing Must determine the primary structure, the sequence of amino acids X-ray Crystallography Used to determine the complex tertiary and quaternary structures. 分子必須形成結晶。 NMR (核磁共振)--分析較複雜。 4.10 Proteomics A new scientific discipline dedicated to understanding the complex relationship of disease and protein expression Uses protein microarrays to test variation in protein expression between healthy and disease states 各跑2D電泳，比對兩者間的差異，取出定序。 補充： monoclonal antibody製造 免疫小鼠，取血清為polyclonal antibody。特異性較低，不可重複製造。 取免疫小鼠之脾細胞，與瘤細胞融合，經篩選得不死的細胞，能分泌單株抗體。特異性高，可重複大量製造，特性完全相同。 * 台大莊榮輝教授網站：.tw/ECX/monoclonal.htm * * 單源抗體的應用 酵素免疫分析ELISA, Western blot 親和性管柱或磁珠快速分離濃縮 flow cytometry immunohistochemistry therapeuticmagic bullet 接藥物或ADCC(Ab dependent cell-mediatedd cytotoxicity)殺死癌細胞 中和細胞激素，如Denosumab中和RANKL，抑制osteoclast成熟，半年打一針防骨質疏鬆 目前在臨床測試的生物醫藥30%為單株抗體，市場300億美金/年 * * * Copyright ? 2009 Pearson Education, Inc. Copyright ? 2009 Pearson Education, Inc. PowerPoint Lectures for Introduction to Biotechnology, Second Edition William J.Thieman and Michael A.Palladino Lectures by Lara Dowland Chapter 4 Proteins as Products Monoclonal antibody as an example(補充) 吳輔祐教授 生物技術與動物科學系 Chapter Contents 4.1 Introduction to Proteins as Biotech Products 4.2 Proteins as Biotechnology Products 4.3 Protein Structures 4.4 Protein Production 4.5 Protein Purification Methods 4.6 Verification 4.7 Preserving Proteins 4.8 Scale-Up of Protein Purificatio