Development of phage biopanning strategies to identify affinity peptide ligands for kappa light chain Fab fragments

Nascimento André; Mullerpatan Akshat; Azevedo Ana Margarida; Karande Pankaj; Cramer Steven

摘要

Abstract >In this work, two phage biopanning strategies were developed to identify affinity peptides for a single Fab and multiple kappa Fabs. For the biopanning rounds, protein L beads were employed to bind Fab targets in a fixed orientation, and NHS functionalized magnetic beads were used to facilitate evaluation of low pH elution conditions. The resulting peptide sequences were synthesized and the binding to different Fabs was evaluated using fluorescence polarization. The first biopanning approach yielded a peptide with similar affinities for two forms of the Fab (recombinantly expressed and post papain‐digestion) as well as the intact antibody. While moderate affinity was observed toward a murine variant of the Fab with the same complementarity determining regions (CDR) region but different framework, minimal binding occurred to a Fab with high sequence homology but containing different CDR loops. The second biopanning strategy yielded a peptide with affinity for all three kappa Fabs indicating that it may be a good lead for the development of more general affinity reagents for recombinant kappa Fabs. Finally, an affinity peptide column was developed, and its efficacy was demonstrated for Fab purification from a complex cell culture fluid mixture. The results presented in this article demonstrate that different peptide‐based phage biopanning strategies can be effectively employed to identify affinity peptide leads for specific Fab and more general kappa Fab purifications. </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> 在这项工作中，开发了两种噬菌体生物丙酸策略以鉴定单个Fab和多个κBabs的亲和肽。对于生物丙烯，使用蛋白质L珠粒在固定取向中结合Fab靶，NHS官能化磁珠用于促进低pH洗脱条件的评估。合成所得肽序列，使用荧光极化评价与不同FAB的结合。第一种生物丙宁方法得到了具有类似亲和力的肽，用于两种形式的Fab（重组表达和后蛋白质消化）以及完整的抗体。朝着具有相同互补性确定区域（CDR）区域但不同骨架的鼠鼠变体观察到中等亲和力，但具有高序列同源物的Fab，但含有不同的CDR环，而且含有不同的CDR环，而且含有不同的CDR环，但朝向具有相同的互补性确定区域（CDR）区域的鼠变体。第二种生物丙策略产生了对所有三种κB晶叶草的亲和力的肽，表明它可能是用于制备重组κBabs的更一般亲和试剂的良好铅。最后，开发了亲和肽柱，并证明了其从复合细胞培养流体混合物中纯化的疗效。本文提出的结果表明，可以有效地利用不同的基于肽的噬菌体生物杀策略，以鉴定特定Fab的亲和肽引线，更一般的Kappa Fab纯化。</ 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6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=Nascimento André&option=202" target="_blank" rel="nofollow">Nascimento André;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Mullerpatan Akshat&option=202" target="_blank" rel="nofollow">Mullerpatan Akshat;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Azevedo Ana Margarida&option=202" target="_blank" rel="nofollow">Azevedo Ana Margarida;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Karande Pankaj&option=202" target="_blank" rel="nofollow">Karande Pankaj;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Cramer Steven&option=202" target="_blank" rel="nofollow">Cramer Steven;</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>iBB – Institute for Bioengineering and Biosciences Instituto Superior TécnicoUniversidade de LisboaLisbon Portugal;</p> <p>Department of Chemical and Biological Engineering Center for Biotechnology and Interdisciplinary StudiesRensselaer Polytechnic InstituteTroy New York;</p> <p>iBB – Institute for Bioengineering and Biosciences Instituto Superior TécnicoUniversidade de LisboaLisbon Portugal;</p> <p>Department of Chemical and Biological Engineering Center for Biotechnology and Interdisciplinary StudiesRensselaer Polytechnic InstituteTroy New York;</p> <p>Department of Chemical and Biological Engineering Center for Biotechnology and Interdisciplinary StudiesRensselaer 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=affinity chromatography&option=203" rel="nofollow">affinity chromatography;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=affinity peptides&option=203" rel="nofollow">affinity peptides;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Fab fragments&option=203" rel="nofollow">Fab fragments;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=phage display&option=203" rel="nofollow">phage display;</a> </p> <div class="translation"> 机译：亲和层析;亲和肽;Fab碎片;噬菌体展示; 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href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=赵明军&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,赵明军</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=邬向东&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,邬向东</a> <span> <a href="/conference-cn-28970/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 中国微生物学会兽医微生物专委会学术年会中国畜牧兽医学会生物制品学分会第九次学术研讨会 </a> <span> <span> . 2003</span> </span> </div> </li> <li> <div> <b>7. </b><a class="enjiyixqcontent" href="/academic-degree-domestic_mphd_thesis/020311312800.html">抗汉坦病毒单克隆抗体可变区基因分析及其噬菌体配体肽的筛选和鉴定</a> <b>[A] </b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=王长军&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 王长军</a> <span> . 2003</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/06120106566592.html">蛋白质中赖氨酸丁烯酰化的鉴定方法及其亲和试剂研制</a> <b>[P]</b> . <span> 中国专利： CN102375065A </span> <span> . 2012-03-14</span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/06120106280624.html">利用噬菌体抗体库针对人血浆蛋白组的亲和配体筛选的方法</a> <b>[P]</b> . <span> 中国专利： CN102095871A </span> <span> . 2011-06-15</span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130435343851.html">light chain, heavy chain, immunoglobulin, humanized antibody. pharmaceutical and therapeutic composition, isolated polypeptide and peptide, polypeptide variant, isolated nucleic acid molecule, vector, host cell, transgenic animal, use of the variable region sequence presented as sequence id: 2 or sequence id. 4 and methods of preventing or treating an amyloidogenic disease associated with amyloid deposition of fir trees in a patient's brain and alzheimer's disease, of producing an antibody or fragment thereof, of identifying residues sensitive to substitution in a variable frame region of immunoglobulin , imaging of amyloid deposits in a patient's brain and reducing plaque, neuter and beta amyloid peptide (fir) load and neuritically dystrophy</a> <b>[P]</b> . <span> 外国专利：  BR0308143A </span> <span> . 2007-01-23</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：轻链，重链，免疫球蛋白，人源化抗体。药物和治疗组合物，分离的多肽和肽，多肽变体，分离的核酸分子，载体，宿主细胞，转基因动物，可变区序列的使用，其以序列ID：2或序列ID表示。 4和预防或治疗与患者大脑中的冷杉树的淀粉样蛋白沉积有关的淀粉样变性疾病和阿尔茨海默氏病的方法，产生抗体或其片段，鉴定对免疫球蛋白可变框架区中的取代敏感的残基，淀粉样蛋白成像的方法沉积在患者的大脑中并减少斑块，中性和β淀粉样肽（fir）负荷和神经营养不良 </span> </p> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/patent-detail/06130408641653.html">method for making product chain for benchmarking of product development strategy of corporation using big data appratus for making product chain for benchmarking of product development strategy of corporation using big data and storage medium for storing a program for making product chain for benchmarking of product development strategy of corporation using big data</a> <b>[P]</b> . <span> 外国专利：  KR101693014B1 </span> <span> . 2017-01-05</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：大数据的企业产品开发战略基准的产品链制作方法，大数据的企业产品开发战略基准的产品链制作方法和存储产品开发战略基准的产品链存储程序的存储介质公司使用大数据 </span> </p> </li> <li> <div> <b>5. </b><a class="enjiyixqcontent" href="/patent-detail/06130411505817.html">METHOD FOR MAKING PRODUCT CHAIN FOR BENCHMARKING OF PRODUCT DEVELOPMENT STRATEGY OF CORPORATION USING BIG DATA, APPRATUS FOR MAKING PRODUCT CHAIN FOR BENCHMARKING OF PRODUCT DEVELOPMENT STRATEGY OF CORPORATION USING BIG DATA, AND STORAGE MEDIUM FOR STORING A PROGRAM FOR MAKING PRODUCT CHAIN FOR BENCHMARKING OF PRODUCT DEVELOPMENT STRATEGY OF CORPORATION USING BIG DATA</a> <b>[P]</b> . <span> 外国专利：  KR20160051264A </span> <span> . 2016-05-11</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：使用大数据对公司的产品开发策略进行产品链标记的方法，使用大数据对公司的产品开发策略进行产品链标记的装置以及用于对产品进行销售的产品链进行存储的程序的存储介质大数据的公司战略 </span> </p> </li> </ul> </div> </div> </div> <div class="theme cardcommon" style="overflow: auto;display:none"> <h3 class="all_title" id="enpatent55">相关主题</h3> <ul id="subject"> </ul> </div> </div> </div> </div> <div class="right rightcon"> <div class="details_img cardcommon clearfix" style="margin-bottom: 10px;display:none;" > </div> </div> </div> <div id="thesis_get_original1" class="downloadBth" style="bottom: 19px;z-index: 999;" onclick="ywcd('0704021880454','4',7,2,1,'',this,24)" class="delivery" prompt="010401" title="通过人工服务将文献原文发送至邮箱" >获取原文</div> <div class="journalsub-pop-up" style="display: none"> <div class="journal-sub"> <h2>期刊订阅</h2> <img 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