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首页> 外文期刊>Biotechnology and Bioengineering >Understanding of decreased sialylation of Fc‐fusion protein in hyperosmotic recombinant Chinese hamster ovary cell culture: NN ‐glycosylation gene expression and NN ‐linked glycan antennary profile
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Understanding of decreased sialylation of Fc‐fusion protein in hyperosmotic recombinant Chinese hamster ovary cell culture: NN ‐glycosylation gene expression and NN ‐linked glycan antennary profile

机译：对高血症重组中FC融合蛋白的降低唾液酸化唾液酸蛋白的理解： n n-glycosylation基因表达和< i xmlns =“http://www.wiley.com/namespaces/wiley”> n n -linked glycan antiennary配置文件

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ABSTRACT <section xml:id="bit26284-sec-0001"> > To understand the effects of hyperosmolality on protein glycosylation, recombinant Chinese hamster ovary (rCHO) cells producing the Fc‐fusion protein were cultivated in hyperosmolar medium resulting from adding NaCl (415?mOsm/kg). The hyperosmotic culture showed increased specific Fc‐fusion protein productivity ( q <sub>Fc</sub> ) but a decreased proportion of acidic isoforms and sialic acid content of the Fc‐fusion protein. The intracellular and extracellular sialidase activities in the hyperosmotic cultures were similar to those in the control culture (314?mOsm/kg), indicating that reduced sialylation of Fc‐fusion protein at hyperosmolality was not due to elevated sialidase activity. Expression of 52 N ‐glycosylation‐related genes was assessed by the NanoString nCounter system, which provides a direct digital readout using custom‐designed color‐coded probes. After 3 days of hyperosmotic culture, nine genes ( ugp , slc35a3 , slc35d2 , gcs1 , manea , mgat2 , mgat5b , b4galt3 , and b4galt4 ) were differentially expressed over 1.5‐fold of the control, and all these genes were down‐regulated. N ‐linked glycan analysis by anion exchange and hydrophilic interaction HPLC showed that the proportion of highly sialylated (di‐, tri‐, tetra‐) and tetra‐antennary N ‐linked glycans was significantly decreased upon hyperosmotic culture. Addition of betaine, an osmoprotectant, to the hyperosmotic culture significantly increased the proportion of highly sialylated and tetra‐antennary N ‐linked glycans ( P? ≤?0.05), while it increased the expression of the N ‐glycan branching/antennary genes ( mgat2 and mgat4b ). Thus, decreased expression of the genes with roles in the N ‐glycan biosynthesis pathway correlated with reduced sialic acid content of Fc‐fusion protein caused by hyperosmolar conditions. Taken together, the results obtained in this study provide a better understanding of the detrimental effects of hyperosmolality on N ‐glycosylation, especially sialylation, in rCHO cells. Biotechnol. Bioeng. 2017;114: 1721–1732. ? 2017 Wiley Periodicals, Inc. </section> </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 XMLNS =“http://www.wiley.com/namespaces/wiley”type =“main”xml：lang =“en”> <title type =“main”>抽象</ title> <section XML：ID =“Bit26284-SEC-0001”> >理解高氧化术对蛋白质糖基化的影响，在加入NaCl（415Ω/ mOSm /公斤）。高骨质培养表现出较高的特异性Fc-融合蛋白质生产率（ Q </ i> <sub> Fc </ sub>），但是酸性同种型的比例降低和Fc-融合蛋白的唾液酸含量。高骨质培养物中的细胞内和细胞外酶活性与对照培养物（314〜MOSM / kg）的细胞外酶活性相似，表明在高氧化酶的高氧态溶液中降低了Fc融合蛋白的唾液酸化不是由于唾液酸酶活性升高。通过纳米管道系统评估52 <I> N </ I> -Glycosylation相关基因的表达，该系统提供了使用定制设计的颜色编码探针的直接数字读数。经过3天的高骨肉培养，九个基因（ UGP </ i>， SLC35A3，<I> SLC35D2，<I> GCS1 </ I>，甘肃</ i>， MgAT2，<I> MgAT5b </ i>， B4gALT3 </ i>和 B4gALT4 </ I>）差异地表达超过1.5倍对照，所有这些基因都被下调。阴离子交换和亲水性相互作用HPLC的<I> N </ I> - 链接的聚糖分析表明，高唾液酸化（二，三 - ，四 - ）和四天数的比例</ i> - 链接高骨肉培养物显着降低了聚糖。添加甜菜碱，一种渗透培养物，对高蛋白培养物显着增加了高度唾液酸化和四抗长期的比例 -Lind聚糖（p≤β/ i>≤？0.05）增加了 n </ i> - 甘露甘露糖分支/抗长基因的表达（ mgat2和 mgat4b）。因此，与由高硅基条件引起的Fc融合蛋白的降低的唾液酸含量相关的 n〜甘露甘油生物合成途径中的基因的表达降低。在一起，本研究中获得的结果提供了更好地了解高氧化溶液对RCHO细胞中的高氧化溶液的不利影响，特别是唾液酸化。 Biotechnol。生物。 2017; 114：1721-1732。还2017 Wiley期刊，Inc。</ p> </ section> </ abstract> </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-15017/'>《Biotechnology and Bioengineering》</a> <b style="margin: 0 2px;">|</b><span>2017年第8期</span><b style="margin: 0 2px;">|</b><span>共12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=Lee Jong Hyun&option=202" target="_blank" rel="nofollow">Lee Jong Hyun;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Jeong Yeong Ran&option=202" target="_blank" rel="nofollow">Jeong Yeong Ran;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Kim Yeon‐Gu&option=202" target="_blank" rel="nofollow">Kim Yeon‐Gu;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Lee Gyun Min&option=202" target="_blank" rel="nofollow">Lee Gyun Min;</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>Department of Biological SciencesKAIST 335 Gwahak‐ro Yuseong‐gu Daejeon 305‐701 Republic of Korea;</p> <p>Department of Biological SciencesKAIST 335 Gwahak‐ro Yuseong‐gu Daejeon 305‐701 Republic of Korea;</p> <p>Biotechnology Process Engineering CenterKRIBB Ochang‐eup Cheongwon‐gu Cheongju Republic of Korea;</p> <p>Department of Biological SciencesKAIST 335 Gwahak‐ro Yuseong‐gu Daejeon 305‐701 Republic of Korea;</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/180.html" title="生物工程学（生物技术）">生物工程学（生物技术）;</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=betaine&option=203" rel="nofollow">betaine;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=CHO cells&option=203" rel="nofollow">CHO cells;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Fc‐fusion protein&option=203" rel="nofollow">Fc‐fusion protein;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=hyperosmolality&option=203" rel="nofollow">hyperosmolality;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=nanostring nCounter system&option=203" rel="nofollow">nanostring nCounter system;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=N ‐glycosylation&option=203" rel="nofollow">N ‐glycosylation;</a> </p> <div class="translation"> 机译：甜菜碱;CHO细胞;FC-融合蛋白;高氧化性;纳米杆菌系统;n-glycosylation; </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 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