Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 411, Issue 18, Pages 4193-4202Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-019-01825-y
Keywords
ICPMS; Quantification; Nanodrug; Internalization; CD47-peptide; SIRP alpha
Funding
- National Natural Science Foundation of China [21535007, 21874112, 21475108]
- National Basic Research 973 Program [2014CB932004]
- National Science and Technology Basic Work [2015FY111400]
- Foundation for Innovative Research Groups of the National Natural Science Foundation of China [21521004]
- Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT) [IRT13036]
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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) (Ru-102) versus traditional laser confocal scanning microscope (lambda (ex/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) x 10(4) Ru(bpy)(3)@SiO2-COOH and (0.28 +/- 0.04) x 10(4) 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 alpha (SIRP alpha) on the macrophage by measuring the clickable tagged Eu using ICPMS. Furthermore, the interaction between CD47-peptide and SIRP alpha as well as the changes of the remaining free SIRP alpha during the internalization process of Ru(bpy)(3)@SiO2@CD47-peptide NPs were quantitatively evaluated, providing direct experimental evidence of the longspeculated crucial CD47-SIRP alpha 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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