Monitoring in Vivo Behaviors of Protein Nanocages via Encapsulating an NIR-II Ag2S Quantum Dot

Chunyan Li , Feng Li , Yejun Zhang , Wenjing Zhang , Xian-En Zhang , Qiangbin Wang
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Abstract

Introduction

Protein nanocages (PNCs) have been recognized as a promising platform for nanomedicine innovation. Real-time in vivo tracking of PNCs can provide critically important information for the development of PNC-based diagnostics and therapeutics. Here we demonstrate a strategy for monitoring the behaviors of PNCs in vivo by encapsulating a Ag2S quantum dot (QD) with fluorescence in the second near-infrared window (NIR-II, 1000-1700 nm) inside the PNC, using a simian virus 40 (SV40) PNC (PNCSV40) as a model.

Methods

The Ag2S QD was encapsulated into the PNCSV40 through controllable molecular self-assembly. The dynamic migration of Ag2S@PNCSV40 in living mouse was tracked in real time under an InGaAs-based shortwave infrared imaging system and was further corroborated by ex vivo imaging, inductively coupled plasma mass spectrometry analysis, and macrophage endocytosis assay.

Results

Benefitting from the high spatiotemporal resolution and deep tissue penetration of NIR-II fluorescence imaging, the dynamic distribution of the PNCSV40 in living mice was tracked in real time with high fidelity, revealing rapid clearance from bloodstream within 5 min post-intravenous injection and selective accumulation in liver, spleen and bone marrow. Furthermore, adopting the PEGylation strategy, PEGylated PNCSV40 presents remarkably different behaviors in vivo with significantly prolonged blood circulation and much less uptake in the reticuloendothelial system (RES), leading to desirable pharmacokinetics and pharmacodynamics of PNC-based nanomedicines.

Discussion

This study represents the first evidence of real-time tracking of the intrinsic behaviors of PNCs in vivo without interference in PNC-host interactions by encapsulating nanoprobes inside, instead of conjugating nanoprobes onto the outer surface of PNCs. The as-described imaging strategy will facilitate the study of interactions between exogenously introduced PNCs and host body, prompting the development of future protein-based drugs, high-efficacy targeted delivery system, sensors, etc.

通过包裹NIR-II Ag2S量子点监测蛋白质纳米笼在体内的行为
蛋白质纳米笼(pnc)已被认为是纳米医学创新的一个有前途的平台。pnc的实时体内跟踪可以为基于pnc的诊断和治疗的发展提供至关重要的信息。本文以猿猴病毒40 (SV40) PNC (PNCSV40)为模型,通过在PNC内的第二个近红外窗口(NIR-II, 1000-1700 nm)封装荧光Ag2S量子点(QD)来监测PNC在体内的行为。方法通过可控的分子自组装将Ag2S QD包封到PNCSV40中。利用基于ingaas的短波红外成像系统实时跟踪Ag2S@PNCSV40在活体小鼠体内的动态迁移,并通过离体成像、电感耦合血浆质谱分析和巨噬细胞内吞实验进一步证实。结果利用NIR-II荧光成像的高时空分辨率和深组织穿透性,实时、高保真地跟踪了PNCSV40在活体小鼠体内的动态分布,揭示了PNCSV40在静脉注射后5 min内迅速从血流中清除,并在肝脏、脾脏和骨髓中选择性积累。此外,采用聚乙二醇化策略,聚乙二醇化的PNCSV40在体内表现出显著不同的行为,血液循环明显延长,网状内皮系统(RES)的摄取大大减少,导致pnc纳米药物具有理想的药代动力学和药效学效果。本研究首次证明了在不干扰pnc与宿主相互作用的情况下,通过将纳米探针封装在pnc内部,而不是将纳米探针偶联到pnc的外表面,可以实时跟踪pnc在体内的内在行为。所述成像策略将有助于研究外源引入的pnc与宿主体之间的相互作用,促进未来基于蛋白质的药物、高效靶向递送系统、传感器等的发展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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