Zhang Genwei, Brown Joseph S, Quartararo Anthony J, Li Chengxi, Tan Xuyu, Hanna Stephanie, Antilla Sarah, Cowfer Amanda E, Loas Andrei, Pentelute Bradley L
Massachusetts Institute of Technology, Department of Chemistry, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA.
FogPharma, 30 Acorn Park Dr, Cambridge, MA, 02140, USA.
Commun Chem. 2022 Jan 19;5(1):8. doi: 10.1038/s42004-022-00625-3.
Rapid discovery and development of serum-stable, selective, and high affinity peptide-based binders to protein targets are challenging. Angiotensin converting enzyme 2 (ACE2) has recently been identified as a cardiovascular disease biomarker and the primary receptor utilized by the severe acute respiratory syndrome coronavirus 2. In this study, we report the discovery of high affinity peptidomimetic binders to ACE2 via affinity selection-mass spectrometry (AS-MS). Multiple high affinity ACE2-binding peptides (ABP) were identified by selection from canonical and noncanonical peptidomimetic libraries containing 200 million members (dissociation constant, K = 19-123 nM). The most potent noncanonical ACE2 peptide binder, ABP N1 (K = 19 nM), showed enhanced serum stability in comparison with the most potent canonical binder, ABP C7 (K = 26 nM). Picomolar to low nanomolar ACE2 concentrations in human serum were detected selectively using ABP N1 in an enzyme-linked immunosorbent assay. The discovery of serum-stable noncanonical peptidomimetics like ABP N1 from a single-pass selection demonstrates the utility of advanced AS-MS for accelerated development of affinity reagents to protein targets.
快速发现并开发出对蛋白质靶点具有血清稳定性、选择性和高亲和力的肽基结合物具有挑战性。血管紧张素转换酶2（ACE2）最近被确定为一种心血管疾病生物标志物，也是严重急性呼吸综合征冠状病毒2利用的主要受体。在本研究中，我们报告了通过亲和选择质谱法（AS-MS）发现对ACE2具有高亲和力的拟肽结合物。通过从包含2亿个成员的经典和非经典拟肽文库中进行筛选，鉴定出多种高亲和力的ACE2结合肽（ABP）（解离常数K = 19 - 123 nM）。与最有效的经典结合肽ABP C7（K = 26 nM）相比，最有效的非经典ACE2肽结合物ABP N1（K = 19 nM）在血清中表现出更高的稳定性。在酶联免疫吸附测定中，使用ABP N1可选择性地检测人血清中皮摩尔至低纳摩尔浓度的ACE2。从单次筛选中发现像ABP N1这样具有血清稳定性的非经典拟肽，证明了先进的AS-MS在加速开发针对蛋白质靶点的亲和试剂方面的实用性。
