生物工程下游技术-绪论

单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,Downstream Processing of Bioengineering,生物工程下游技术,杨志红,yangzhihong,691612,Section One,Preface,Considerable effort and time is allocated to introducing cell culture and fermentation technology to undergraduate students in academia, generally through a range of courses in industrial biotechnology and related disciplines. Similarly, a large number of textbooks are available to describe the applications of these technologies in industry.,However, there has been a general lack of appreciation of the significant developments in downstream processing and isolation technology, the need for which is largely driven by the stringent regulatory requirements for purity and quality of,injectable,biopharmaceuticals. This is particularly reflected by the general absence of coverage of this subject in many biotechnology and related courses in educational institutions.,For a considerable while I have felt that there is increasing need for an introductory text to various aspects of downstream processing, particularly with respect to the needs of the biopharmaceutical and biotechnology industry. Although there are numerous texts that cover various aspects of protein purification techniques in isolation, there is a need for a work that covers the broad range of isolation technology in an industrial setting.,It is anticipated that,Downstream Processing of Proteins: Methods and Protocols,will play a small part in filling this gap and thus prove a useful contribution to the field. It is also designed to encourage educational strategists to broaden the coverage of these topics in industrial biotechnology courses by including accounts of this important and rapidly developing element of the industrial process.,The hope is that this will result in graduates having a reasonable understanding of downstream processing principles and techniques, and thus be better prepared to fulfill the ever-increasing demand for competent isolation scientists in industries.,This is, of course, achieved with the help of the dedicated contributing authors of Downstream Processing of Proteins: Methods and Protocols, without whose willingness to contribute and patience it would not have been possible. I would also like to thank the Humana Press and Prof. John Walker (the series editor) for their encouragement and prompt feedback.,My thanks are also due to the,Medeva,Pharma,Development management for providing me with the time and opportunity to fulfill this task, and without whose support it would have been impossible. Finally, I wish to thank my wife, children, and family members for allowing me to persevere with my editing activities in perhaps what should have been their time.,Section Two,菌种筛选,摇瓶实验,发酵罐实验,Industrial Scale up,To transfer the pilot scale results into a commercially feasible production,setting.,Fermentor,sizes range from 100 L to 500,000 L, depending on products.,在疾病治疗方面的应用,生物制药,抗生素,基因治疗,生物工程研究领域,微生物培养,动物细胞培养,植物细胞培养,天然资源,生化材料,海洋生物培养,生物工程产业领域,生物制药,（抗生素，基因重组蛋白，氨基酸，疫苗，菌苗，天然药物，生化药物，血液制品，抗体，多糖，多肽）,生物化工,（乳酸，柠檬酸，苹果酸，丙烯酸，甘油，异丙醇，乙烯）,生物能源,（甲醇，乙醇，生物柴油，生物汽油）,生物材料,（明胶，胶原蛋白，人造皮肤，人造骨，人造脏器）,生物医学,（诊断试剂，基因治疗，人及生物克隆）,环境生物,（环境治理，水污染，土壤污染，风沙治理）,生物食品,（醋，啤酒业，酿酒，乳制品，奶制品）,生物资源,（动物，植物，微生物）,生物农业,（基因食品，基因植物）,生物工程的上中下游,上游：,菌种，基因工程，分子生物学，遗传学,中游：,微生物发酵工程，动植物细胞， 海洋生物培养,下游：,生物分离工程,生物产品特点,产物浓度低的水溶液,原因：,a.,氧传递限制；,b.,细胞量,;c.,产物抑制,组分复杂,a.,大分子,;b.,小分子,;c.,可溶物,;d.,不可溶物,;e.,化学添加物,产物稳定性差,a.,化学降解,(pH ,温度,);,b.,微生物降解 （酶作用， 染菌）,质量要求高 （药品或食品）,生物分离工艺要求,(1),高纯度,(2),大规模,(3),经济性,(4),生物相容性,生物过程开发目标,(1),产品在最短时间内进入市场,(2),符合所有安全要求,(3),工艺成本适当,(4),可靠,(5),安全（放大期间安全尤为重要）,质量控制意义,(1),来自管理机构的压力,(2),公众的觉醒,(3),市场扩张与竞争,(4),安全,(5),提高专一性作用，减少负作用,非蛋白类杂质的去除,（,1,）,DNA,A.,阴离子交换（,pH 4.0),B.,亲和层析（不被吸附）,C.,疏水层析,（,2,）热原 （蛋白质溶液中的去热原）,A.,生产过程无菌,B.,所有层析介质无菌,C.,所用溶液无菌,D.,亲和层析（多粘菌素）,（,3,）去病毒,A.,加热 （,60,),B.,过滤,C.,灭活剂,目标蛋白的表征和分析方法,（,1,）,HPLC,（,2,）,SDS-PAGE,（,3,） 氨基酸顺序分析,（,4,） 氨基酸，肽图，免疫化学,（,5,） 与标准品对照分析,工业应用的生物分离技术,回收技术,:,絮凝，离心，过滤，微过滤。,细胞破碎技术,:,球磨，高压匀浆，化学破碎技术。,初步纯化技术,:,盐析法，有机溶剂沉淀，化学沉淀，大孔吸附树剂，膜分离技术。,高度纯化技术,:,各类层析，亲和，疏水，聚焦，离子交换。,成品加工,-,喷雾干燥,:,气流干燥，沸腾干燥，冷冻干燥，结晶。,生物技术产品的类型,按分子量大小,小分子产品,：（小于,1000,）抗生素，有机酸，氨基酸,大分子产品,：（大于,1000,）酶,抗体，多肽，蛋白质,按产品所处的位置,细胞内,:,胰岛素，干扰素，重组蛋白质,细胞外,：抗生素，胞外酶,下游加工过程的沿革,传统产业（第一代）,19,世纪,60,年代,-20,世纪,50,年代,酒精，丙酮，丁醇,第二代生物技术产品,20,世纪,40,年代,抗生素，有机酸，核酸，酶制剂，单细胞蛋白,第三代,20,世纪,70,年代中期,动物细胞培养,植物细胞培养,基因工程发酵产品,生物下游加工过程的选择准则,步聚少,次序合理,产品规格 （注射，非注射）,生产规模,物料组成,产品形式 （固体,-,适当结晶， 液体,-,适当浓缩）,产品稳定性,物性 （溶解度， 分子电荷，分子大小，功能团，稳定性，挥发性）,危害性,废水处理,生物技术下游加工过程的发展动向,基础理论研究,A,选择性分离剂,B,数学模型,应用研究,A,新老技术的深化研究与融合,B,下游技术与上游技术相结合,(,藕合分离）,C,强化化学作用对分离能力的影响,D,改进上游因素（改进菌种，培养基与发酵条件）,工程问题研究,改善环境相容性,现代生物分离过程的研究方向与特点,（,1,）大规模，高选择性,（液膜萃取，两水相萃取，反渗透，纳米过滤，渗透蒸发，亲和膜过滤，亲和错流过滤， 亲和沉淀 ，分子筛，分子蒸馏）,（,2,）集成化,（扩张床，两水相，液膜分离）,（,3,）快速分离,（色谱）,（,4,）极端条件下分离,（超临界，超声波，超重力）,（,5,）在位分离,（边发酵边分离）,（,6,）环境相容,（超临界）,（,7,）可再生循环,（亲和沉淀，可再生两水相）,Separation Processes:,Extractors,溶质在二个互不相容的溶剂的分配系数不同，所造成的溶质的转移。,混合器和澄清器,膜技术,微滤,超滤,纳滤,反渗透,悬浮粒子,大分子,糖，二价盐,游离酸,单价盐,不游离酸,水,膜的分类与特征,分子印迹分离,超临界流体分离,Phase separation,Top phase,Bottom phase,Recycle,Light,pH,Light,pH,Recycling Polymers Forming Aqueous Two-phase Systems,亲和沉淀,lower critical solution temperature,LCST,生物分离技术与化工分离技术的区别,化工分离技术：,获得纯的化学物质。,生物分离技术：,在得到纯的生物物质同时，还必须关注特定杂质的去除。,与传统的化学试剂的纯度概念不同，生物产物对,有害物质,有严格的控制，生产,过程,也要求有严格的管理，在最终产品中往往不允许有极微量的有害杂质存在。,生物分离技术的重要性,生物产物的特殊性；,生物产物所处环境的复杂性；,对生物产品要求的严格性；,最终结果：,导致下游加工过程中成本往往占整个生物加工过程生产成本的大部分。,下游加工技术的一般流程,生物下游加工过程是指目标产物的分离纯化过程，包括,产物提取,(isolation),产物浓缩,(concentration),产物纯化,(purification),成品化,(polishing),注意,：,多步分离导致收率降低；,分离技术的选择依据,产物所处的位置；,产物性质（分子大小、疏水性、电荷形式和溶解度等）；,生物加工过程自身的规模和产品的商业价值；一种目标产物的分离手段往往不止一种，根据生产的规模和价值，选择合适的分离技术,生物下游加工过程的特点,满足,维持,生物物质,活性,的要求,满足,快速分离,的要求,满足纯度和,杂质去除,的要求,满足,高效分离,的要求,满足,成本优化,的要求,生物分离本质,有效地识别混合物中不同溶质间,物理、化学和生物学性质,的差别，利用能够识别这些差别的分离介质和（或）扩大这些差别的分离设备实现溶质间的分离或目标组分的纯化。,常用的分离技术及其机理,物理性质,力学性质,重力、离心力、筛分,热力学性质,状态变化、相平衡,传质性质,粘度、扩散、热扩散,电磁性质,电泳、电渗、磁化,化学性质,化学热力学,化学平衡,反应动力学,反应速率,光化学性质,激光激发、离子化,生物学性质,分子识别,生物亲和作用、生物学识别,输送性质,生物膜输送,反应、响应、控制,免疫系统,Question,1,、,please introduce the downstream processing of biotechnology in your words ( at least 150 words in,english,).,