Effect of linkers on immobilization of scFvs with biotin–streptavidin interaction

机译：接头对生物素 - 链霉抗生物素蛋白相互作用的SCFV固定的影响

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Abstract > Single‐chain variable fragment antibodies (scFvs) are attractive for use in applications that require high specificity and binding to a target, such as biosensors. Previously, we demonstrated that a variety of scFvs can be immobilized onto a streptavidin surface through in vivo biotinylation of the biotin carboxyl carrier protein (BCCP) or smaller AviTag fused to the scFvs. However, the BCCP constructs showed better immobilization than the AviTag constructs. In this work, we investigated whether the discrepancy between the biotinylation tags could be alleviated by incorporating a flexible (G <sub>4</sub> S) <sub>n</sub> linker of varying lengths or a rigid (EA <sub>3</sub> K) <sub>3</sub> linker between the biotinylation tags and the scFvs scFv13R4 and scFv5. Fusion of the (G <sub>4</sub> S) <sub>5</sub> linker or the (G <sub>4</sub> S) <sub>3</sub> linker to the AviTag construct of scFv13R4 or scFv5, respectively, and fusion of the (EA <sub>3</sub> K) <sub>3</sub> linkers to the AviTag constructs of both scFvs enhanced immobilization. Meanwhile, the robust immobilization of the BCCP construct of the scFv constructs remained unaffected. The positive to neutral effects of the linkers, with no adverse effects, make them beneficial tools to incorporate into fusion proteins that show poor immobilization without a linker. </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> >单个 - 链可变片段抗体（SCFV）是有吸引力的，适用于需要高特异性和结合靶的应用，例如生物传感器。以前，我们证明，各种SCVS可以通过在体内羧基载体蛋白（BCCP）的生物素羧基载体蛋白（BCCP）的生物素化或融合于SCFV的较小横向的生物素化。然而，BCCP构建体显示比Avitag构建体更好地固定。在这项工作中，我们研究了通过掺入柔性（G <sub> 4 </ sub> s） <sub> n </ sub> </ i>的改变的链接器来缓解生物素化标签之间的差异吗？长度或刚性（EA <sub> 3 </ sub> k）<sub> 3 </ sub>链接器之间的生物素化标签和scfvs scfv13r4和scfv5之间。（g <sub> 4 </ sub> s）<sub> 5 </ sub>链接器或（g <sub> 4 </ sub> s）<sub> 3 </ sub>链接器到avitag分别构建SCFV13R4或SCFV5，以及（EA <sub> 3 </ sub> k）<sub> 3 </ sub>接头的融合到SCFV的AVITAG构建体，增强了固定化。同时，SCFV构建体的BCCP构建体的稳健固定仍未受到影响。阳性与接头的中性效果，没有不良反应，使其成为掺入融合蛋白的有益工具，该蛋白质没有接头而显示出差的固定。 </ p> </ 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-15016/'>《Biotechnology and Applied Biochemistry》</a> <b style="margin: 0 2px;">|</b><span>2018年第4期</span><b style="margin: 0 2px;">|</b><span>共6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=Ikonomova Svetlana P.&option=202" target="_blank" rel="nofollow">Ikonomova Svetlana P.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Le Megan T.&option=202" target="_blank" rel="nofollow">Le Megan T.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Kalla Neha&option=202" target="_blank" rel="nofollow">Kalla Neha;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Karlsson Amy J.&option=202" target="_blank" rel="nofollow">Karlsson Amy J.;</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 Biomolecular EngineeringUniversity of MarylandCollege Park MD USA;</p> <p>Department of Chemical and Biomolecular EngineeringUniversity of MarylandCollege Park MD USA;</p> <p>Department of Chemical and Biomolecular EngineeringUniversity of MarylandCollege Park MD USA;</p> <p>Department of Chemical and Biomolecular EngineeringUniversity of MarylandCollege Park MD USA;</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><a href="https://www.zhangqiaokeyan.com/clc/177.html" title="生物化学">生物化学;</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=biotinylation tags&option=203" rel="nofollow">biotinylation tags;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=flexible&option=203" rel="nofollow">flexible;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=rigid&option=203" rel="nofollow">rigid;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=immobilization&option=203" rel="nofollow">immobilization;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=peptide linkers&option=203" rel="nofollow">peptide linkers;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=scFv&option=203" rel="nofollow">scFv;</a> </p> <div class="translation"> 机译：生物素化标签;柔性;刚性;固定化;肽接头;SCFV; </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 >中文文献</li> <li >专利</li> </ul> </div> <div class="similarity_details"> <ul > <li> <div> <b>1. </b><a class="enjiyixqcontent" 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</div> </li> <li> <div> <b>6. </b><a class="enjiyixqcontent" href="/academic-conference-cn_meeting-36950_thesis/020221533427.html">生物素－抗生物素蛋白体系标记ＤＮＡ探针</a> <b>[C]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=陈俊杰&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 陈俊杰</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=游乐然&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,游乐然</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=王若菡&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,王若菡</a> <span> <a href="/conference-cn-36950/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 全国免疫标记技术讨论会 </a> <span> <span> . 1989</span> </span> </div> </li> <li> <div> <b>7. </b><a class="enjiyixqcontent" href="/academic-degree-domestic_mphd_thesis/020313456464.html">乳头状甲状腺癌与结节性甲状腺肿组织内源性抗生物素蛋白结合活性的不同及其临床意义</a> <b>[A] </b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=廉莹&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 廉莹</a> <span> . 2013</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/06120114041099.html">生物素-链霉抗生物素蛋白裂解组合物和文库片段裂解</a> <b>[P]</b> . <span> 中国专利： CN114144523A </span> <span> . 2022-03-04</span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/061201286475.html">低免疫原性链霉抗生物素蛋白及其应用</a> <b>[P]</b> . <span> 中国专利： CN102325884B </span> <span> . 2013.11.20</span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130454399239.html">MODIFIED MOLECULE OF THE AVIDIN TYPE BINDING BIOTIN, ITS USE, NITRO-TYROSINE NATURAL AVIDIN AND PROCESS OF ITS PREPARATION, BACTERIAL STREPTAVIDIN, COLUMN FOR IMMOBILIZATION OF BIOTINYLATED LIGAND AND PROCESS OF OBTAINING AVIDIN COLUMN OR THE BIOTINYLATED LIGAND</a> <b>[P]</b> . <span> 外国专利：  CZ401697A3 </span> <span> . 1998-07-15</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：亲和素型生物素的修饰分子，其用途，硝基酪氨酸天然亲和素及其制备方法，细菌链霉抗生物素，用于固定化生物素化的配体的柱子以及获得的亲和柱或生物素的过程 </span> </p> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/patent-detail/06130454394164.html">MODIFIED MOLECULE OF THE AVIDIN TYPE BINDING BIOTIN, ITS USE, NITRO-TYROSINE NATURAL AVIDIN AND PROCESS OF ITS PREPARATION, BACTERIAL STREPTAVIDIN, COLUMN FOR IMMOBILIZATION OF BIOTINYLATED LIGAND AND PROCESS OF OBTAINING AVIDIN COLUMN OR THE BIOTINYLATED LIGAND</a> <b>[P]</b> . <span> 外国专利：  CZ9704016A3 </span> <span> . 1998-07-15</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：亲和素型生物素的修饰分子，其用途，硝基酪氨酸天然亲和素及其制备方法，细菌链霉抗生物素，用于固定化生物素化的配体的柱子以及获得的亲和柱或生物素的过程 </span> </p> </li> <li> <div> <b>5. </b><a class="enjiyixqcontent" href="/patent-detail/06130430581925.html">STREPTAVIDIN-BIOTIN-LINK ANTIBODY-HAPTEN SYSTEM</a> <b>[P]</b> . <span> 外国专利：  US2009093618A1 </span> <span> . 2009-04-09</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：链霉亲和素-生物素-链接抗体半抗原系统 </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;" 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