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cGMP production and analysis of BG505 SOSIP.664, an extensively glycosylated, trimeric HIV‐1 envelope glycoprotein vaccine candidate

机译：CGMP生产和分析BG505 SOSIP.664，一种广泛的糖基化，三聚体HIV-1包络糖蛋白疫苗候选

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Abstract <section xml:id="bit26498-sec-0001" numbered="no"> > We describe the properties of BG505 SOSIP.664 HIV‐1 envelope glycoprotein trimers produced under current Good Manufacturing Practice (cGMP) conditions. These proteins are the first of a new generation of native‐like trimers that are the basis for many structure‐guided immunogen development programs aimed at devising how to induce broadly neutralizing antibodies (bNAbs) to HIV‐1 by vaccination. The successful translation of this prototype demonstrates the feasibility of producing similar immunogens on an appropriate scale and of an acceptable quality for Phase I experimental medicine clinical trials. BG505 SOSIP.664 trimers are extensively glycosylated, contain numerous disulfide bonds and require proteolytic cleavage, all properties that pose a substantial challenge to cGMP production. Our strategy involved creating a stable CHO cell line that was adapted to serum‐free culture conditions to produce envelope glycoproteins. The trimers were then purified by chromatographic methods using a 2G12 bNAb affinity column and size‐exclusion chromatography. The chosen procedures allowed any adventitious viruses to be cleared from the final product to the required extent of 12 log <sub>10</sub> . The final cGMP production run yielded 3.52?g (peptidic mass) of fully purified trimers (Drug Substance) from a 200?L bioreactor, a notable yield for such a complex glycoprotein. The purified trimers were fully native‐like as judged by negative‐stain electron microscopy, and were stable over a multi‐month period at room temperature or below and for at least 1 week at 50 ° C. Their antigenicity, disulfide bond patterns, and glycan composition were consistent with trimers produced on a research laboratory scale. The meth </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 =“Bit26498-SEC-0001”编号=“否”> >我们描述了在当前良好的制造实践（CGMP）条件下生产的BG505 SOSIP.664 HIV-1封闭糖蛋白三种三蛋白三蛋白三蛋白的特性。这些蛋白质是一种新一代的天然三聚体，是许多结构引导的免疫原性发育计划的基础，其旨在通过疫苗接种促进如何诱导促使截面抗体（BNAB）到HIV-1的基础。该原型的成功翻译证明了在适当的规模和可接受的I期实验医学临床试验中生产类似免疫因素的可行性。 BG505 SOSIP.664三聚体是广泛的糖基化，含有许多二硫键，需要蛋白水解裂解，所有的性质对CGMP生产构成了大量挑战。我们的策略涉及创造一种稳定的CHO细胞系，适应无血清培养条件以产生包膜糖蛋白。然后使用2G12BNAB亲和柱和尺寸排阻色谱法通过色谱法纯化三聚体。所选择的程序允许从最终产品中清除任何不偶诱导病毒，以＆ 12 log <sub> 10 </ sub>。最终的CGMP生产运行产生3.52μg（肽质量）的全纯化的三聚体（药物物质），来自200·L生物反应器，对这种复合糖蛋白的显着产率产生了显着的产率。纯化的三聚体是通过阴性染色电子显微镜判断的完全海底，并且在室温或低于室温下的多月份，并且在50℃/℃下稳定，至少1周。它们的抗原性，二硫键模式和聚糖组合物与在研究实验室规模上产生的三聚体一致。 meth </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>2018年第4期</span><b style="margin: 0 2px;">|</b><span>共15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=Dey Antu K.&option=202" target="_blank" rel="nofollow">Dey Antu K.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Cupo Albert&option=202" target="_blank" rel="nofollow">Cupo Albert;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ozorowski Gabriel&option=202" target="_blank" rel="nofollow">Ozorowski Gabriel;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Sharma Vaneet K.&option=202" target="_blank" rel="nofollow">Sharma Vaneet K.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Behrens Anna‐Janina&option=202" target="_blank" rel="nofollow">Behrens Anna‐Janina;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Go Eden P.&option=202" target="_blank" rel="nofollow">Go Eden P.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ketas Thomas J.&option=202" target="_blank" rel="nofollow">Ketas Thomas J.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Yasmeen Anila&option=202" target="_blank" rel="nofollow">Yasmeen Anila;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Klasse Per J.&option=202" target="_blank" rel="nofollow">Klasse Per J.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Sayeed Eddy&option=202" target="_blank" rel="nofollow">Sayeed Eddy;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Desaire Heather&option=202" target="_blank" rel="nofollow">Desaire Heather;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Crispin Max&option=202" target="_blank" rel="nofollow">Crispin Max;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Wilson Ian A.&option=202" target="_blank" rel="nofollow">Wilson Ian A.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Sanders Rogier W.&option=202" target="_blank" rel="nofollow">Sanders Rogier W.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Hassell Thomas&option=202" target="_blank" rel="nofollow">Hassell Thomas;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ward Andrew B.&option=202" target="_blank" rel="nofollow">Ward Andrew B.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Moore John P.&option=202" target="_blank" rel="nofollow">Moore John P.;</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>International AIDS Vaccine InitiativeNew York New York;</p> <p>Department of Microbiology and ImmunologyWeill Medical College of Cornell UniversityNew York New York;</p> <p>Department of Integrative Structural and Computational Biology International AIDS Vaccine Initiative (IAVI) Neutralizing Antibody Center and the Collaboration for AIDS Vaccine DiscoveryThe Scripps Research InstituteLa Jolla California;</p> <p>International AIDS Vaccine InitiativeNew York New York;</p> <p>Department of Biochemistry Oxford Glycobiology InstituteUniversity of OxfordOxford UK;</p> <p>Department of ChemistryThe University of KansasLawrence Kansas;</p> <p>Department of Microbiology and ImmunologyWeill Medical College of Cornell UniversityNew York New York;</p> <p>Department of Microbiology and ImmunologyWeill Medical College of Cornell UniversityNew York New York;</p> <p>Department of Microbiology and ImmunologyWeill Medical College of Cornell UniversityNew York New York;</p> <p>International AIDS Vaccine InitiativeNew York New York;</p> <p>Department of ChemistryThe University of KansasLawrence Kansas;</p> <p>Department of Biochemistry Oxford Glycobiology InstituteUniversity of OxfordOxford UK;</p> <p>Department of Integrative Structural and Computational Biology International AIDS Vaccine Initiative (IAVI) Neutralizing Antibody Center and the Collaboration for AIDS Vaccine DiscoveryThe Scripps Research InstituteLa Jolla California;</p> <p>Department of Microbiology and ImmunologyWeill Medical College of Cornell UniversityNew York New York;</p> <p>International AIDS Vaccine InitiativeNew York New York;</p> <p>Department of Integrative Structural and Computational Biology International AIDS Vaccine Initiative (IAVI) Neutralizing Antibody Center and the Collaboration for AIDS Vaccine DiscoveryThe Scripps Research InstituteLa Jolla California;</p> <p>Department of Microbiology and ImmunologyWeill Medical College of Cornell UniversityNew York New York;</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=affinity purification&option=203" rel="nofollow">affinity purification;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=cGMP&option=203" rel="nofollow">cGMP;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=HIV‐1 vaccine&option=203" rel="nofollow">HIV‐1 vaccine;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=native‐like Env trimers&option=203" rel="nofollow">native‐like Env trimers;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=recombinant vaccine development&option=203" rel="nofollow">recombinant vaccine development;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=SOSIP&option=203" rel="nofollow">SOSIP;</a> </p> <div class="translation"> 机译：亲和力纯化;CGMP;HIV-1疫苗;天然型ENVINER;重组疫苗开发;索普; </div> </li> </ul> </div> </div> <div class="literature cardcommon"> <div class="similarity "> <h3 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</span> <span> . 2006</span><span>,第1期</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/061203602296.html">一种高效快速的多糖蛋白结合疫苗纯化分析方法</a> <b>[P]</b> . <span> 中国专利： CN106397537B </span> <span> . 2020.01.07</span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/06120108881982.html">一种高效快速的多糖蛋白结合疫苗纯化分析方法</a> <b>[P]</b> . <span> 中国专利： CN106397537A </span> <span> . 2017-02-15</span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130433116637.html">Hiv-1 Neutralizing Antibodies Elicited By Trimeric Hiv-1 Envelope Glycoprotein Complex</a> <b>[P]</b> . <span> 外国专利： <!-- 美国专利： --> US2008274134A1 </span> <span> . 2008-11-06</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：三聚体Hiv-1包膜糖蛋白复合物产生的Hiv-1中和抗体 </span> </p> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/patent-detail/06130432179638.html">HIV-1 NEUTRALIZING ANTIBODIES ELICITED BY TRIMERIC HIV-1 ENVELOPE GLYCOPROTEIN COMPLEX</a> <b>[P]</b> . <span> 外国专利： <!-- 欧洲知识产权局专利： --> EP1766097A4 </span> <span> . 2008-03-19</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：三元HIV-1包膜糖蛋白复合物引发的HIV-1中和抗体 </span> </p> </li> <li> <div> <b>5. </b><a class="enjiyixqcontent" href="/patent-detail/06130434592646.html">HIV-1 NEUTRALIZING ANTIBODIES ELICITED BY TRIMERIC HIV-1 ENVELOPE GLYCOPROTEIN COMPLEX</a> <b>[P]</b> . <span> 外国专利： <!-- 欧洲知识产权局专利： --> EP1766097A2 </span> <span> . 2007-03-28</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：三元HIV-1包膜糖蛋白复合物引发的HIV-1中和抗体 </span> </p> </li> </ul> </div> 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