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Fluorescent and mass spectrometric evaluation of the phagocytic internalization of a CD47-peptide modified drug-nanocarrier

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Abstract

Ru(bpy)3@SiO2-COOH and Ru(bpy)3@SiO2@CD47-peptide nanoparticles (NPs) with fluorescent and mass spectrometric properties were designed and synthesized as the models of drug-nanocarriers. Their phagocytic internalization could be quantitatively measured using more sensitive inductively coupled plasma mass spectrometry (ICPMS) (102Ru) versus traditional laser confocal scanning microscope (λex/em = 458/600 nm) for the first time. Modification of a self-signal trigging CD47-peptide on the NPs’ surface decreased internalization by 10 times, (2.79 ± 0.21) × 104 Ru(bpy)3@SiO2-COOH and (0.28 ± 0.04) × 104 Ru(bpy)3@SiO2@CD47-peptide NPs per RAW264.7 macrophage (n = 5). The alkynyl-linked CD47-peptide allowed us to quantify the number (2412 ± 250) of CD47-peptide modified on the NP and the total content (5.14 ± 0.25 amol) of signal regulatory protein α (SIRPα) on the macrophage by measuring the clickable tagged Eu using ICPMS. Furthermore, the interaction between CD47-peptide and SIRPα as well as the changes of the remaining free SIRPα during the internalization process of Ru(bpy)3@SiO2@CD47-peptide NPs were quantitatively evaluated, providing direct experimental evidence of the longspeculated crucial CD47-SIRPα interaction for drug-nanocarriers to escape internalization by phagocytic cells. Remarkable difference in the internalization ratio of 12.3 ± 4.8 of Ru(bpy)3@SiO2-COOH NPs and 4.3 ± 0.5 Ru(bpy)3@SiO2@CD47-peptide NPs with and without the protein corona indicated that CD47-peptide still worked when the protein corona formed. Not limited to the evaluation of the NPs studied here, such a fluorescent and mass spectrometric approach is very much expected to apply to the assessment of other drug-nanocarriers designed by chemists and before their medical applications.

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Acknowledgements

We thank the financial supports from the National Natural Science Foundation of China (21535007, 21874112 and 21475108), the National Basic Research 973 Program (2014CB932004) and the National Science and Technology Basic Work (2015FY111400) as well as the Foundation for Innovative Research Groups of the National Natural Science Foundation of China (21521004) and Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT, Grant IRT13036).

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  1. Department of Chemistry & the Key Laboratory of Spectrochemical Analysis and Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, Fujian, China

    Chunlan Liu, Danxia Yu, Fuchun Ge, Limin Yang & Qiuquan Wang

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  1. Chunlan Liu
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  2. Danxia Yu
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Correspondence to Qiuquan Wang.

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Published in the topical collection New Insights into Analytical Science in China with guest editors Lihua Zhang, Hua Cui, and Qiankun Zhuang.

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Liu, C., Yu, D., Ge, F. et al. Fluorescent and mass spectrometric evaluation of the phagocytic internalization of a CD47-peptide modified drug-nanocarrier. Anal Bioanal Chem 411, 4193–4202 (2019). https://doi.org/10.1007/s00216-019-01825-y

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