A novel scale‐down mimic of perfusion cell culture using sedimentation in an automated microbioreactor (SAM)

Kreye Steffen; Stahn Rainer; Nawrath Karina; Goralczyk Vicky; Zoro Barney; Goletz Steffen

摘要

Abstract > Continuous upstream processing in mammalian cell culture for recombinant protein production holds promise to increase product yield and quality. To facilitate the design and optimization of large‐scale perfusion cultures, suitable scale‐down mimics are needed which allow high‐throughput experiments to be performed with minimal raw material requirements. Automated microbioreactors are available that mimic batch and fed‐batch processes effectively but these have not yet been adapted for perfusion cell culture. This article describes how an automated microbioreactor system (ambr15) can be used to scale‐down perfusion cell cultures using cell sedimentation as the method for cell retention. The approach accurately predicts the viable cell concentration, in the range of about 1 × 10 7 ?cells/mL for a human cell line, and cell viability of larger scale cultures using a hollow fiber based cell retention system. While it was found to underpredict cell line productivity, the method accurately predicts product quality attributes, including glycosylation profiles, from cultures performed in bioreactors with working volumes between 1 L and 1,000?L. The spent media exchange method using the ambr15 was found to predict the influence of different media formulations on large‐scale perfusion cultures in contrast to batch and chemostat experiments performed in the microbioreactor system. The described experimental setup in the microbioreactor allowed an 80‐fold reduction in cell culture media requirements, half the daily operator time, which can translate into a cost reduction of approximately 2.5‐fold compared to a similar experimental setup at bench scale. </abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> <div class="translation abstracttxt"> <span class="zhankaihshouqi fivelineshidden" id="abstract"> <span>机译：</span><Abstract Type =“Main”XML：Lang =“en”> <标题类型=“main”>抽象</ title> > 哺乳动物细胞培养中的连续上游加工用于重组蛋白质产量的承担能力增加产品产量和质量。为了便于大规模灌注培养的设计和优化，需要合适的缩放模拟，从而允许以最小的原材料要求进行高通量实验。自动微生物反应器可有效地使用模拟批量和补料分批过程，但这些尚未适用于灌注细胞培养物。本文介绍了如何使用细胞沉积作为细胞保留方法的灌注细胞培养物如何使用自动微生物反应器系统（AMBR15）。该方法准确地预测可行的细胞浓度，在约1×10的范围内 7 </ sup> ？使用中空纤维基细胞保留系统，使用中空纤维的细胞系细胞的细胞/ ml为人体细胞系的细胞活力。虽然发现弱势细胞系高生产率，但该方法精确地预测产品质量属性，包括糖基化型材，从生物反应器中进行的培养物，其工作量在1L和1,000？L之间进行。发现使用AMBR15的废媒体交换方法预测与微生物反应器系统中进行的批量和化疗实验相比，不同介质制剂对大规模灌注培养物的影响。微生物反应器中所描述的实验设置允许降低细胞培养介质要求的80倍，每日操作员时间的一半，与工作台规模的类似实验设置相比，可以转化为约2.5倍的成本降低。 </ p> </摘要> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> </div> <div class="record"> <h2 class="all_title" id="enpatent33" >著录项</h2> <ul> <li> <span class="lefttit">来源</span> <div style="width: 86%;vertical-align: text-top;display: inline-block;"> <a href='/journal-foreign-15005/'>《Biotechnology Progress》</a> <b style="margin: 0 2px;">|</b><span>2019年第5期</span><b style="margin: 0 2px;">|</b><span>共11页</span> </div> </li> <li> <div class="author"> <span class="lefttit">作者</span> <p id="fAuthorthree" class="threelineshidden zhankaihshouqi"> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Kreye Steffen&option=202" target="_blank" rel="nofollow">Kreye Steffen;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Stahn Rainer&option=202" target="_blank" rel="nofollow">Stahn Rainer;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Nawrath Karina&option=202" target="_blank" rel="nofollow">Nawrath Karina;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Goralczyk Vicky&option=202" target="_blank" rel="nofollow">Goralczyk Vicky;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zoro Barney&option=202" target="_blank" rel="nofollow">Zoro Barney;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Goletz Steffen&option=202" target="_blank" rel="nofollow">Goletz Steffen;</a> </p> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zkzz" style="display: none;">展开▼</span> </div> </li> <li> <div style="display: flex;"> <span class="lefttit">作者单位</span> <div style="position: relative;margin-left: 3px;max-width: 639px;"> <div class="threelineshidden zhankaihshouqi" id="fOrgthree"> <p>Research and Development Glycotope GmbHBerlin Germany;</p> <p>Research and Development Glycotope GmbHBerlin Germany;</p> <p>Research and Development Glycotope GmbHBerlin Germany;</p> <p>Research and Development Glycotope GmbHBerlin Germany;</p> <p>Sartorius Stedim BiotechRoyston UK;</p> <p>Department of Biotechnology and BiomedicineTechnical University of DenmarkLyngby Denmark;</p> </div> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zhdw" style="display: none;">展开▼</span> </div> </div> </li> <li > <span class="lefttit">收录信息</span> <span style="width: 86%;vertical-align: text-top;display: inline-block;"></span> </li> <li> <span class="lefttit">原文格式</span> <span>PDF</span> </li> <li> <span class="lefttit">正文语种</span> <span>eng</span> </li> <li> <span class="lefttit">中图分类</span> <span><a href="https://www.zhangqiaokeyan.com/clc/15.html" title="生物科学">生物科学;</a></span> </li> <li class="antistop"> <span class="lefttit">关键词</span> <p style="width: 86%;vertical-align: text-top;"> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=cell culture&option=203" rel="nofollow">cell culture;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=high throughput&option=203" rel="nofollow">high throughput;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=microbioreactor&option=203" rel="nofollow">microbioreactor;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=perfusion&option=203" rel="nofollow">perfusion;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=scale‐down&option=203" rel="nofollow">scale‐down;</a> </p> <div class="translation"> 机译：细胞培养;高吞吐量;微生物反应器;灌注;缩放; </div> </li> </ul> </div> </div> <div class="literature cardcommon"> <div class="similarity "> <h3 class="all_title" id="enpatent66">相似文献</h3> <div class="similaritytab clearfix"> <ul> <li class="active" 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href="/ntis-science-report_ad_thesis/0207168901.html">Nonyloxytryptamine Mimics Polysialic Acid and Modulates Neuronal and Glial Functions in Cell Culture.</a> <b>[R] </b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Loers, G.&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Loers, G.,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Saini, V.&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Saini, V.,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Mishra, B.&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Mishra, B.,</a> <span>2014</span> </span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：Nonyloxytryptamine模拟聚唾液酸并调节细胞培养中的神经元和胶质细胞功能。</span> </p> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/academic-journal-cn_academic-abstracts-china-colleges-universities-mechanical-engineering_thesis/0201288859306.html">Dimensional synthesis of a novel 5-DOF reconfigurable hybrid perfusion manipulator for large-scale spherical honeycomb perfusion</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Hui YANG&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . Hui YANG</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Hairong FANG&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Hairong FANG</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Yuefa FANG&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Yuefa FANG</a> <span> <a href="/journal-cn-8555/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 中国高等学校学术文摘·机械工程 </a> </span> <span> . 2021</span><span>,第001期</span> </span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/academic-conference-cn_meeting-28517_thesis/020222027518.html">Metal-alumina cermet inert anode in a 100A bench-scale cell for aluminium electrolysis</a> <b>[C]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Qiu Zhuxian&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . Qiu Zhuxian</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Shi Zhongning&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Shi Zhongning</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Xu Junli&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Xu Junli</a> <span> <a href="/conference-cn-28517/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 2004年有色金属冶金及材料第二届国际学术会议 </a> <span> <span> . 2004</span> </span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/academic-degree-domestic_mphd_thesis/020315010264.html">perfusion and immunofluorescence staining. Retinal explant culture was used to examine the effect of CNTF, MIF and MSF in the environment macrophages were not readily available</a> <b>[A] </b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=岑令平&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 岑令平</a> <span> . 2006</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/06130501682598.html">A RAMAN SPECTROSCOPY INTEGRATED PERFUSION CELL CULTURE SYSTEM FOR MONITORING AND AUTO-CONTROLLING PERFUSION CELL CULTURE</a> <b>[P]</b> . <span> 外国专利：  EP3976753A1 </span> <span> . 2022-04-06</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：用于监测和自动控制灌注细胞培养的拉曼光谱集成灌注细胞培养系统 </span> </p> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/06130421792057.html">Method for culturing a cell culture in an automated cell culture system and automated cell culture system</a> <b>[P]</b> . <span> 外国专利：  DE102005021034B4 </span> <span> . 2012-09-13</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：在自动化细胞培养系统中培养细胞培养物的方法和自动化细胞培养系统 </span> </p> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130434151210.html">Method for culturing a cell culture in an automated cell culture system and automated cell culture system</a> <b>[P]</b> . <span> 外国专利：  DE102005021034A1 </span> <span> . 2006-11-16</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：在自动化细胞培养系统中培养细胞培养物的方法和自动化细胞培养系统 </span> </p> </li> </ul> </div> </div> </div> <div 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