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介质原理→关键性能→评估→哪些评价的控制策略→收集数据评价→结果→结论
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构建特性→下游纯化工艺
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裂解为有菌区，后续粗纯为无活菌区（工程菌）
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验证策略控制与工艺设计相关
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属性：分子筛？官能团？
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产品工艺和工艺功能
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功能原理结合蛋白性质来写
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大分子分子筛效应不明显，扩散弱
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柱效：均一性、松紧：对称性，N代表分离能力
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分子筛
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全排阻，N不重要，对成型重要，类似与化工类的蒸馏塔
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脱盐、除色素
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非全排阻（进孔）
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路径不一样
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反压大→碎胶多→装柱效果差→筛胶
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日常维护（装填、封存）
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清洁（前后处理、CIP）
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# 来源类型
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天然
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表达
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# 踏板
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引用化工蒸馏的概念
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# 装柱工艺
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# 柱效测定
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小分子扩散探测评价介质分布情况
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图谱（吸附类的）：产品来评价吸附，官能团多少直接影响吸附能力和分离能力
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柱效
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定期测柱效的标准
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合格限
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行动限
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警戒限
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中间柱效变化由图谱体现，如果中间某次图谱异常，怎么办？监测下样液合格即可放行？
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上样液可以过滤，减少柱子的压力，延长层析柱的使用寿命
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柱头膜堵塞
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反向冲洗、超声波冲洗，目视看不出来
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柱壳问题，正向上样不行，可以尝试反向上样
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缓冲液中，EDTA（金属螯合剂，抢夺酶的金属离子）抑制酶的作用
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# 工艺参数
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柱桶规格
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柱高
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柱内直径
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径高比
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上样载量（体积）：上样体积大，滞留时间长
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上样浓度：浓度高，热扩散明显，蛋白的溶解上限
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样品状态：粘度、颗粒物
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基线：规定基线范围
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柱效要求
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线性流速：流速高，介质耐受问题；流速低，滞留时间长，分离度差
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缓冲体系
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柱温（以流出液温度为准）和环境温度
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# 洗脱
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分离度：连续梯度洗脱优于恒定洗脱
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恒定洗脱
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连续梯度洗脱
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手工较难实现，一般通过设备实现
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# 图谱
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根据图谱可以反推柱子和样品的状态
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