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Implementation of an experimental and computational tool set to study protein‐mAb interactions

机译：实施实验和计算工具，用于研究蛋白质-MAb相互作用

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Abstract >This work focused on the development of a combined experimental and computational tool set to study protein‐mAb interactions. A model protein library was first screened using cross interaction chromatography to identify proteins with the strongest retention. Fluorescence polarization was then employed to study the interactions and thermodynamics of the selected proteins—lactoferrin, pyruvate kinase, and ribonuclease B with the mAb. Binding affinities of lactoferrin and pyruvate kinase to the mAb were seen to be relatively salt insensitive in the range examined. Further, a strong entropic contribution was observed, suggesting the importance of hydrophobic interactions. On the other hand, ribonuclease B‐mAb binding was seen to be enthalpically driven and salt sensitive, indicating the importance of electrostatic interactions. Protein–protein docking was then carried out and the results identified the CDR region on the mAb as an important binding site for all three proteins. The binding interfaces identified for the mAb‐lactoferrin and mAb‐pyruvate kinase systems were found to contain complementary hydrophobic and oppositely charged clusters on the interacting regions which were indicative of both hydrophobic and electrostatic interactions. On the other hand, the binding site on ribonuclease B was predominantly positively charged with minimal hydrophobicity. This resulted in an alignment with negatively charged clusters on the mAb, supporting the contention that these interactions were primarily electrostatic in nature. Importantly, these computational results were found to be consistent with the fluorescence polarization data and this combined approach may have utility in examining mAb‐HCP interactions which can often complicate the downstream processing of biologics. ? 2019 American Institute of Chemical Engineers </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> >这项工作的重点是开发组合的实验和计算工具，以研究蛋白质-MAb相互作用。首先使用交叉相互作用色谱法筛选模型蛋白质文库，以鉴定具有最强保留的蛋白质。然后采用荧光偏振来研究选定的蛋白质 - 乳铁蛋白，丙酮酸激酶和核糖核酸酶B的相互作用和热力学与mAb。在检查范围内，乳铁蛋白和丙酮酸激酶的结合亲和力与MAb相对盐不敏感。此外，观察到强烈的熵贡献，表明疏水性相互作用的重要性。另一方面，核糖核酸酶B-MAB结合被认为是焓驱动和盐敏感，表明静电相互作用的重要性。然后进行蛋白质 - 蛋白酶对接，结果将MAB上的CDR区域鉴定为所有三种蛋白质的重要结合位点。发现针对MAB-LACTOFERIN和MAB-丙酮酸激酶体系鉴定的结合界面在相互作用区域上含有互补的疏水性和相对的带电簇，其指示疏水性和静电相互作用。另一方面，Ribonuclease B上的结合位点主要被带正电荷，疏水性最小。这导致MAB上的带负电荷簇的对准，支持这些相互作用主要静电的争夺。重要的是，发现这些计算结果与荧光偏振数据一致，并且这种组合方法可以在检查MAB-HCP相互作用时具有效用，这通常会使生物制剂的下游加工复杂化。还2019年美国化学工程师学院</ 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4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=Ranjan Swarnim&option=202" target="_blank" rel="nofollow">Ranjan Swarnim;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Chung Wai Keen&option=202" target="_blank" rel="nofollow">Chung Wai Keen;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zhu Min&option=202" target="_blank" rel="nofollow">Zhu Min;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Robbins David&option=202" target="_blank" rel="nofollow">Robbins David;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Cramer Steven M.&option=202" target="_blank" rel="nofollow">Cramer Steven M.;</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 Chemical and Biological EngineeringRensselaer Polytechnic InstituteTroy New York;</p> <p>Purification Process SciencesMedImmune LLCGaithersburg Maryland;</p> <p>Purification Process SciencesMedImmune LLCGaithersburg Maryland;</p> <p>Purification Process SciencesMedImmune LLCGaithersburg Maryland;</p> <p>Department of Chemical and Biological EngineeringRensselaer Polytechnic InstituteTroy 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/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=docking&option=203" rel="nofollow">docking;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=downstream bioprocessing&option=203" rel="nofollow">downstream bioprocessing;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=fluorescence&option=203" rel="nofollow">fluorescence;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=host cell protein&option=203" rel="nofollow">host cell protein;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=monoclonal antibody&option=203" rel="nofollow">monoclonal antibody;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=protein–protein interaction&option=203" rel="nofollow">protein–protein interaction;</a> </p> <div class="translation"> 机译：对接;下游生物加工;荧光;宿主细胞蛋白;单克隆抗体;蛋白质 - 蛋白质相互作用; </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" >外文文献</li> <li 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Osmari Vendrame,</a> <span>2020</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：靶向β-嵌体的药物 - 药物与纤维蛋白原血浆蛋白质的相互作用：计算和实验研究</span> </p> </li> <li> <div> <b>10. </b><a class="enjiyixqcontent" href="/open-access_resources_thesis/0100024484175.html">Computational Tools and Databases for the Study and Characterization of Protein Interactions.</a> <b>[O] </b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Segura Joan&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Segura Joan,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Blas Pastor Jose Ramon&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Blas Pastor Jose Ramon,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Fernandez-Fuentes Narcis&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Fernandez-Fuentes Narcis </a> <span>2012</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：研究和表征蛋白质相互作用的计算工具和数据库。</span> </p> </li> <li> <div> <b>11. </b><a class="enjiyixqcontent" href="/ntis-science-report_de_thesis/02071742188.html">Computational tools for experimental determination and theoretical prediction of protein structure</a> <b>[R] </b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=O'Donoghue, S. &option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">O'Donoghue, S. ,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Rost, B.&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Rost, B. </a> <span>1995</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：用于蛋白质结构的实验测定和理论预测的计算工具</span> </p> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/academic-journal-cn_science-china-information-sciences_thesis/0201227387363.html">The open-close mechanism of M2 channel protein in influenza A virus:A computational study on the hydrogen bonds and cation-π interactions among His37 and Trp41</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . </a> <span> <a href="/journal-cn-17558/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 中国科学 </a> </span> <span> . 2008</span><span>,第008期</span> </span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/academic-journal-cn_detail_thesis/0201292574553.html">Fluorescence as a Tool to Study Lipid-Protein Interactions: The Case of α-Synuclein</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Azucena Gonzalez-Horta&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . Azucena Gonzalez-Horta</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Brenda Gonzalez Hernandez&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Brenda Gonzalez Hernandez</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Abelardo Chavez-Montes&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Abelardo Chavez-Montes</a> <span> <a href="/journal-cn-56595/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 生物物理学期刊（英文） </a> </span> <span> . 2013</span><span>,第1期</span> </span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/academic-conference-cn_meeting-21907_thesis/02022491101.html">Experimental Studies on HA/TCP with Recombinant Human Bone Morphogentic Protein-2 for Ostioporotic Bone Defect Repair</a> <b>[C]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=DUAN Wangpign&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . DUAN Wangpign</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=ZHANG Min&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,ZHANG Min</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=LI Gang&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,LI Gang</a> <span> <a href="/conference-cn-21907/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 2007国际骨质疏松——骨与关节大会 </a> <span> <span> . 2007</span> </span> </div> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/academic-degree-domestic_mphd_thesis/020315285169.html">Revealing Luminescent Detection Mechanism for Indoor Volatile Organic Compounds via Non--covalent Interaction:A Computational study</a> <b>[A] </b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Manzoor Hussain&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . Manzoor Hussain</a> <span> . 2019</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/06130448243210.html">EXPERIMENTAL TEST COMPLEX FOR STUDY AND OPTIMIZATION OF MONORAIL TRANSPORT SYSTEMS; TRACK AND ROLLING STOCK OF EXPERIMENTAL TEST COMPLEX AND METHOD OF STUDY AND OPTIMIZATION OF MONORAIL TRANSPORT SYSTEMS OF EXPERIMENTAL TEST STAND</a> <b>[P]</b> . <span> 外国专利：  RU2167403C1 </span> <span> . 2001-05-20</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：用于研究和优化单轨运输系统的实验测试复合体;实验台架的轨迹和滚动位置以及实验台架单轨运输系统的研究和优化方法 </span> </p> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/06130408392735.html">A method, computer program product, and computational unit for generating a statistical experimental design for a study on a drive system of a motor vehicle</a> <b>[P]</b> . <span> 外国专利：  DE102017109412A1 </span> <span> . 2017-07-06</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：用于生成统计实验设计以研究机动车辆的驱动系统的方法，计算机程序产品和计算单元 </span> </p> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130417603942.html">experimental wind tunnel for agricultural spraying process (1) for the study of the interactions between the effectiveness of pesticides and the risks they represent the environment and public health under conditions of controlled</a> <b>[P]</b> . <span> 外国专利：  TN2013000088A1 </span> <span> . 2014-06-25</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：用于农业喷洒过程的实验风洞（1），用于研究在受控条件下农药的有效性与农药对环境和公众健康的危害之间的相互作用 </span> </p> </li> </ul> </div> </div> </div> <div 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