• 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 >Rational design of viscosity reducing mutants of a monoclonal antibody: Hydrophobic versus electrostatic inter-molecular interactions
【2h】

Rational design of viscosity reducing mutants of a monoclonal antibody: Hydrophobic versus electrostatic inter-molecular interactions

机译：单克隆抗体降粘突变体的合理设计：疏水与静电分子间相互作用

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

High viscosity of monoclonal antibody formulations at concentrations ≥100 mg/mL can impede their development as products suitable for subcutaneous delivery. The effects of hydrophobic and electrostatic intermolecular interactions on the solution behavior of MAB 1, which becomes unacceptably viscous at high concentrations, was studied by testing 5 single point mutants. The mutations were designed to reduce viscosity by disrupting either an aggregation prone region (APR), which also participates in 2 hydrophobic surface patches, or a negatively charged surface patch in the variable region. The disruption of an APR that lies at the interface of light and heavy chain variable domains, VH and VL, via L45K mutation destabilized MAB 1 and abolished antigen binding. However, mutation at the preceding residue (V44K), which also lies in the same APR, increased apparent solubility and reduced viscosity of MAB 1 without sacrificing antigen binding or thermal stability. Neutralizing the negatively charged surface patch (E59Y) also increased apparent solubility and reduced viscosity of MAB 1, but charge reversal at the same position (E59K/R) caused destabilization, decreased solubility and led to difficulties in sample manipulation that precluded their viscosity measurements at high concentrations. Both V44K and E59Y mutations showed similar increase in apparent solubility. However, the viscosity profile of E59Y was considerably better than that of the V44K, providing evidence that inter-molecular interactions in MAB 1 are electrostatically driven. In conclusion, neutralizing negatively charged surface patches may be more beneficial toward reducing viscosity of highly concentrated antibody solutions than charge reversal or aggregation prone motif disruption.
机译：浓度≥100mg / mL的单克隆抗体制剂的高粘度可能会阻碍其发展为适合皮下给药的产品。通过测试5个单点突变体，研究了疏水和静电分子间相互作用对MAB 1溶液行为的影响，该溶液在高浓度下变得不可接受的粘性。突变被设计为通过破坏易于聚集的区域（APR）来降低粘度，该区域也参与2个疏水性表面补丁，或在可变区域中带负电荷的表面补丁。通过L45K突变破坏位于轻链和重链可变域VH和VL界面的APR，使MAB 1不稳定并消除了抗原结合。但是，在前面的残基（V44K）处的突变也位于相同的APR中，在不牺牲抗原结合或热稳定性的情况下，提高了MAB 1的表观溶解度并降低了其粘度。中和带负电荷的表面贴剂（E59Y）也增加了MAB 1的表观溶解度并降低了其粘度，但在同一位置（E59K / R）上的电荷反转会导致不稳定，溶解度降低并导致样品处理困难，从而使其无法进行粘度测量高浓度。 V44K和E59Y突变均显示出相似的表观溶解度增加。但是，E59Y的粘度曲线比V44K的粘度曲线好得多，这提供了MAB 1中分子间相互作用是静电驱动的证据。总而言之，中和带负电荷的表面斑块可能比降低电荷逆转或易于聚集的基序破坏更有利于降低高浓度抗体溶液的粘度。

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