The Shark Strikes Twice: Hypervariable Loop 2 of Shark IgNAR Antibody Variable Domains and Its Potential to Function as an Autonomous Paratope
The Shark Strikes Twice: Hypervariable Loop 2 of Shark IgNAR Antibody Variable Domains and Its Potential to Function as an Autonomous Paratope
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鲨鱼两次袭击：鲨鱼 IgNAR 抗体可变域的高变环 2 及其作为自主互补位的潜力
DOI：
10.1007/s10126-015-9642-z
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发表时间：
2015
影响因子：
3
通讯作者：
Kolmar H
中科院分区：
文献类型：
--
作者：
Zielonka S;Empting M;Könning D;Grzeschik J;Krah S;Becker S;Dickgießer S;Kolmar H
在本研究中，我们设计了 IgNAR 可变域的高变环 2 (HV2)，使其仅促进抗原结合，可能充当自主互补位。为此，使对应于HV2的表面暴露环多样化，并使用酵母表面展示（YSD）作为平台技术通过文库筛选分离IgNAR抗体（vNAR）分子的抗原特异性可变域。使用上皮细胞粘附分子 (EpCAM) 特异性 vNAR 作为起始材料，并对 HV2 中的 9 个残基进行随机化。针对分化簇 3ε (CD3ε) 和人 Fcγ 分离出包含新的 HV2 介导的互补位的靶标特异性克隆，同时保留对 EpCAM 的高亲和力。本质上，我们证明了包含针对给定靶分子的中等亲和力的新互补位可以被设计到 vNAR 支架中，该支架独立于由互补决定区 3 (CDR3) 和 CDR1 组成的原始抗原结合位点发挥作用。
In this present study, we engineered hypervariable loop 2 (HV2) of the IgNAR variable domain in a way that it solely facilitates antigen binding, potentially functioning as an autonomous paratope. For this, the surface-exposed loop corresponding to HV2 was diversified and antigen-specific variable domain of IgNAR antibody (vNAR) molecules were isolated by library screening using yeast surface display (YSD) as platform technology. An epithelial cell adhesion molecule (EpCAM)-specific vNAR was used as starting material, and nine residues in HV2 were randomized. Target-specific clones comprising a new HV2-mediated paratope were isolated against cluster of differentiation 3ε (CD3ε) and human Fcγ while retaining high affinity for EpCAM. Essentially, we demonstrate that a new paratope comprising moderate affinities against a given target molecule can be engineered into the vNAR scaffold that acts independent of the original antigen-binding site, composed of complementarity-determining region 3 (CDR3) and CDR1.
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影响因子：
2.4
作者：
Benatuil, Lorenzo;Perez, Jennifer M.;Hsieh, Chung-Ming
通讯作者：
Hsieh, Chung-Ming
Shark Attack：通过逐步体外亲和力成熟衍生自鲨鱼 vNAR 结构域的高亲和力结合蛋白
影响因子：
4.1
作者：
Zielonka S;Weber N;Becker S;Doerner A;Christmann A;Christmann C;Fritz J;Schäfer E;Steinmann B;Empting M;Ockelmann P;Lierz M;Kolmar H
通讯作者：
Kolmar H
影响因子：
5.6
作者：
Stanfield, Robyn L.;Dooley, Helen;Wilson, Ian A.
通讯作者：
Wilson, Ian A.
DOI：
10.1073/pnas.0508341103
发表时间：
2006-02-07
影响因子：
11.1
作者：
Dooley, H;Stanfield, RL;Flajnik, MF
通讯作者：
Flajnik, MF