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| 重组人小分子抗体ScFv-Fc发酵条件的优化及纯化 | |
| 引用本文： | 王丁丁,苏曼曼,胡丽莉,袁丽颖,颜炜群.重组人小分子抗体ScFv-Fc发酵条件的优化及纯化[J].现代生物医学进展,2011,11(16):3048-3051. |
| 作者姓名： | 王丁丁 苏曼曼 胡丽莉 袁丽颖 颜炜群 |
| 作者单位： | 1. 广东药学院生命科学与生物制药学院 广东广州 510006 2. 吉林大学药学院 吉林长春 130021 3. 吉林大学附属第三幼儿园 吉林长春 130021 |
| 基金项目： | 国家自然科学基金青年基金，吉林大学科学前沿与交叉学科创新创新项目 |
| 摘 要： | 目的:研究重组人小分子抗体ScFv-Fc在毕赤酵母中分泌表达的最佳条件,以及ScFv-Fc的纯化方法。方法:分别从甲醇浓度、pH、诱导时间等方面对毕赤酵母重组菌株产生ScFv-Fc的发酵过程进行了优化;通过硫酸铵沉淀结合protein A亲和层析柱,对ScFv-Fc的纯化方法进行了研究。结果:确定ScFv-Fc在毕赤酵母中分泌表达的最佳条件为:在pH5.2的条件下,以0.5%甲醇诱导72 h。经过protein A亲和层析柱纯化后,ScFv-Fc纯度可达94%以上。结论:确定了ScFv-Fc在毕赤酵母中分泌表达的最佳条件以及纯化方法,为重组抗体分子诊断、治疗试剂的开发以及抗体的人源化奠定了物质基础。 |
| 关 键 词： | ScFv-Fc 表达 毕赤酵母 发酵优化 纯化 |
An optimized Fermentation and Purification Process for a Recombinant Human Antibody ScFv-Fc Fragment Expressed in Pichia Pastoris |
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| WANG Ding-ding,SU Man-man,HU Li-li,YUAN Li-ying,YAN Wei-qun.An optimized Fermentation and Purification Process for a Recombinant Human Antibody ScFv-Fc Fragment Expressed in Pichia Pastoris[J].Progress in Modern Biomedicine,2011,11(16):3048-3051. | |
| Authors: | WANG Ding-ding SU Man-man HU Li-li YUAN Li-ying YAN Wei-qun |
| Institution: | WANG Ding-ding~1,SU Man-man~2,HU Li-li~1,YUAN Li-ying~3,YAN Wei-qun~2 (1 Institute of Life Science and Biological Pharmacy,Guangdong Pharmaceutical university,Guangzhou,Guangdong Province,China,510006,2College of Pharmaceutical,Jilin University,Changchun,Jilin Province,130021,3the 3rd kindergarten affiliated to Jilin university,130021) |
| Abstract: | Objective:To obtain an optimized fermentation and purification process for high-level production of a recombinant human antibody ScFv-Fc fragment expression secreted in Pichia pastoris.Methods:The growth conditions of the transformant strain were optimized in 50 ml conical tubes including pH,methanol concentration and inducing time.The ScFv-Fc was purified using a two-step scheme:ammonium sulfate fractionation,protein A Sepharose.Results:ScFv-Fc production was found to increase with 0.5%(v/v) methanol conce... |
| Keywords: | ScFv-Fc Expression Pichia pastoris Optimized fermentation Purification |
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