How soft corona, the protein corona’s outer layer, contributes to biological identity of nanomaterials is largely because capturing protein composition of the soft corona in situ remains challenging. We herein develop an in situ Fishing method that can monitor the dynamic formation of protein corona on ultra-small chiral Cu₂S nanoparticles (NPs) allowing us to directly separate and identify the corona protein composition. Our method detects spatiotemporal processes in the evolution of hard and soft coronas on chiral NPs, revealing subtle differences in NP − protein interactions even within several minutes. This study highlights the importance of in situ and dynamic analysis of soft/hard corona, provides insights into the role of soft corona in mediating biological responses of NPs, and offers a universal strategy to characterize soft corona to guide the rational design of biomedical nanomaterials.

软电晕（蛋白质电晕的外层）对纳米材料的生物学特性的贡献在很大程度上是因为原位捕获软电晕的蛋白质组成仍然具有挑战性。我们在此开发了一种原位钓鱼方法，可以监测超小手性 Cu _{2上蛋白质电晕的动态形成}S 纳米颗粒 (NPs) 使我们能够直接分离和识别电晕蛋白的组成。我们的方法检测手性 NP 上硬和软电晕演化的时空过程，即使在几分钟内也能揭示 NP - 蛋白质相互作用的细微差异。本研究强调了软/硬电晕原位和动态分析的重要性，提供了对软电晕在介导 NPs 生物反应中的作用的见解，并提供了一种表征软电晕的通用策略，以指导生物医学纳米材料的合理设计。