• 主题
高级搜索  >
历史搜索 清空历史
首页> 美国卫生研究院文献>mAbs >Stability engineering of anti-EGFR scFv antibodies by rational design of a lambda-to-kappa swap of the VL framework using a structure-guided approach
【2h】

Stability engineering of anti-EGFR scFv antibodies by rational design of a lambda-to-kappa swap of the VL framework using a structure-guided approach

机译：通过使用结构指导方法合理设计VL框架的Lambda-kappa交换来设计抗EGFR scFv抗体的稳定性

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务，本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文，但由于OA文献来源多样且变更频繁，仍可能出现获取不到、文献不完整或与标题不符等情况，如果获取不到我们将提供退款服务。请知悉。
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Phage-display technology facilitates rapid selection of antigen-specific single-chain variable fragment (scFv) antibodies from large recombinant libraries. ScFv antibodies, composed of a VH and VL domain, are readily engineered into multimeric formats for the development of diagnostics and targeted therapies. However, the recombinant nature of the selection strategy can result in VH and VL domains with sub-optimal biophysical properties, such as reduced thermodynamic stability and enhanced aggregation propensity, which lead to poor production and limited application. We found that the C10 anti-epidermal growth factor receptor (EGFR) scFv, and its affinity mutant, P2224, exhibit weak production from E. coli. Interestingly, these scFv contain a fusion of lambda3 and lambda1 V-region (LV3 and LV1) genes, most likely the result of a PCR aberration during library construction. To enhance the biophysical properties of these scFvs, we utilized a structure-based approach to replace and redesign the pre-existing framework of the VL domain to one that best pairs with the existing VH. We describe a method to exchange lambda sequences with a more stable kappa3 framework (KV3) within the VL domain that incorporates the original lambda DE-loop. The resulting scFvs, C10KV3_LV1DE and P2224KV3_LV1DE, are more thermodynamically stable and easier to produce from bacterial culture. Additionally, C10KV3_LV1DE and P2224KV3_LV1DE retain binding affinity to EGFR, suggesting that such a dramatic framework swap does not significantly affect scFv binding. We provide here a novel strategy for redesigning the light chain of problematic scFvs to enhance their stability and therapeutic applicability.
机译：噬菌体展示技术有助于从大型重组文库中快速选择抗原特异性单链可变片段（scFv）抗体。由VH和VL结构域组成的ScFv抗体易于工程化为多聚体形式，以开发诊断和靶向疗法。然而，选择策略的重组性质可能导致VH和VL结构域具有次优的生物物理特性，例如降低的热力学稳定性和增强的聚集倾向，从而导致生产不佳和应用受限。我们发现C10抗表皮生长因子受体（EGFR）scFv及其亲和突变体P2224表现出弱的大肠杆菌生产能力。有趣的是，这些scFv包含lambda3和lambda1 V区（LV3和LV1）基因的融合体，这很可能是文库构建过程中PCR异常的结果。为了增强这些scFv的生物物理特性，我们采用了一种基于结构的方法来替换和重新设计VL结构域的现有框架，使其与现有VH最佳配对。我们描述了一种在包含原始lambda DE环的VL域内与更稳定的kappa3框架（KV3）交换lambda序列的方法。所得的scFvs C10KV3_LV1DE和P2224KV3_LV1DE具有更高的热力学稳定性，更易于从细菌培养物中生产。此外，C10KV3_LV1DE和P2224KV3_LV1DE保留了与EGFR的结合亲和力，表明这种戏剧性的框架互换不会显着影响scFv结合。我们在这里提供了一种新的策略，用于重新设计有问题的scFv的轻链，以增强其稳定性和治疗性。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
代理获取

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号