Engineering Next Generation Cyclized Peptide Ligands for Light‐Controlled Capture and Release of Therapeutic Proteins
Engineering Next Generation Cyclized Peptide Ligands for Light‐Controlled Capture and Release of Therapeutic Proteins
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DOI：
10.1002/adfm.202101410
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发表时间：
2021-05
影响因子：
19
通讯作者：
R. Prodromou;Kevin N. Day;Sahand Saberi-Bosari;John D Schneible;Matthew D. Mabe;A. San Miguel;M. Daniele;V. Pozdin;S. Menegatti
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文献类型：
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作者：
R. Prodromou;Kevin N. Day;Sahand Saberi-Bosari;John D Schneible;Matthew D. Mabe;A. San Miguel;M. Daniele;V. Pozdin;S. Menegatti
光亲和吸附剂（即具有光控生物识别功能的配体功能化的半透明基质）代表了纯化不稳定生物制剂的未来技术。在这项研究中，提出了一个由偶氮苯环化肽（ACPs）偶联到半透明多孔珠（ChemMatrix）的光亲和吸附剂原型的框架。该方法结合了计算和实验工具来设计acp，并研究其光控异构化动力学和蛋白质生物识别。首先，介绍了一种模块化设计，用于定制ACP的构象，促进测序，简化顺式/反式异构体及其差异蛋白质结合的计算机建模。然后，报道了一种用于测量化学基质珠上ACPs光异构化动力学的光谱系统；使用该装置，证明了不同光强下的异构化与环化几何形状有关，特别是与硅计算的反式和顺式异构体的能量差有关。此外，本文还提出了一种用于ACP‐ChemMatrix微珠分选的微流控装置，以血管细胞粘附分子1 （VCAM‐1）为靶蛋白，cycloAZOB[GVHAKQHRN‐K*]‐G‐ChemMatrix为模型光亲和吸附剂，选择和验证光亲和配体。所提出的ACPs表现出快速和明确的光控异构化和生物识别。利用光来控制VCAM - 1的吸附和释放，证明了光亲和吸附剂对生化特性对其纯化构成挑战的目标的潜力。
Photo‐affinity adsorbents (i.e., translucent matrices functionalized with ligands featuring light‐controlled biorecognition) represent a futuristic technology for purifying labile biologics. In this study, a framework for prototyping photo‐affinity adsorbents comprising azobenzene‐cyclized peptides (ACPs) conjugated to translucent porous beads (ChemMatrix) is presented. This approach combines computational and experimental tools for designing ACPs and investigating their light‐controlled isomerization kinetics and protein biorecognition. First, a modular design for tailoring ACP's conformation, facilitating sequencing, and streamlining the in silico modeling of cis/trans isomers and their differential protein binding is introduced. Then, a spectroscopic system for measuring the photo‐isomerization kinetics of ACPs on ChemMatrix beads is reported; using this device, it is demonstrated that the isomerization at different light intensities is correlated to the cyclization geometry, specifically the energy difference of trans versus cis isomers as calculated in silico. Also, a microfluidic device for sorting ACP‐ChemMatrix beads to select and validate photo‐affinity ligands using Vascular Cell Adhesion Molecule 1 (VCAM‐1) as target protein and cycloAZOB[GVHAKQHRN‐K*]‐G‐ChemMatrix as model photo‐affinity adsorbent is presented. The proposed ACPs exhibit rapid and defined light‐controlled isomerization and biorecognition. Controlling the adsorption and release of VCAM‐1 using light demonstrates the potential of photo‐affinity adsorbents for targets whose biochemical liability poses challenges to its purification.