Modulation of single-chain antibody affinity with temperature-responsive elastin-like polypeptide linkers
Modulation of single-chain antibody affinity with temperature-responsive elastin-like polypeptide linkers
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DOI：
10.1021/bm0507002
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发表时间：
2006-04-01
影响因子：
6.2
通讯作者：
Yarmush, ML
中科院分区：
文献类型：
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作者：
Megeed, Z;Winters, RM;Yarmush, ML
单链抗体是基因工程构建体，由抗体的V-H和V-L结构域通过柔性肽接头连接组成，通常为（GGGGS）（3）。我们询问用已知经历环境诱导的结构转变的肽替换该柔性接头是否可以导致具有受控结合和释放特征的抗体。为此，我们遗传修饰并产生了一系列具有通用接头序列（VPGXG）（n）的抗荧光素单链抗体，其中n为1.2至3，X为瓦尔或His，以评估接头长度和组成的影响。我们的研究结果表明，含有弹性蛋白样多肽接头的单链抗体在室温下具有与野生型（GGGGS）3相当的平衡亲和力（K-D），但随着温度的升高，结合动力学改变，配体释放更快。这些结果与已知弹性蛋白样多肽随着温度升高而经历的分子有序性增加和收缩一致。使用刺激响应性接头调节抗体亲和力可在生物传感器、药物递送和生物分离中具有应用。
Single-chain antibodies are genetically engineered constructs composed of a V-H and V-L domain of an antibody linked by a flexible peptide linker, commonly (GGGGS)(3). We asked whether replacement of this flexible linker with peptides known to undergo environmentally induced structural transitions could lead to antibodies with controlled binding and release characteristics. To this end, we genetically modified and produced a series of anti-fluorescein single-chain antibodies with the general linker sequence (VPGXG)(n), where n is 1.2 to 3 and X is Val or His, to evaluate the effects of linker length and composition. Our results indicate that single-chain antibodies containing elastin-like polypeptide linkers have equilibrium affinity (K-D) comparable to wild-type (GGGGS)3 at room temperature but altered binding kinetics and faster ligand release as the temperature is raised. These results are consistent with the increased molecular order and contraction that elastin-like polypeptides are known to undergo with increased temperature. Modulation of antibody affinity using stimulus-responsive linkers may have applications in biosensors, drug delivery, and bioseparations.