期刊
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
卷 1858, 期 7, 页码 1499-1506出版社
ELSEVIER
DOI: 10.1016/j.bbamem.2016.03.023
关键词
Drug delivery; Cellular uptake; Internalization pathways; Intracellular distribution; Intracellular degradation; Toxicity threshold
资金
- NYU Abu Dhabi
- NYU Abu Dhabi Institute [G1205]
- National Institutes of Health [F31 AG038110]
Cell-penetrating peptides (CPPs) have emerged as a potentially powerful tool for drug delivery due to their ability to efficiently transport a whole host of biologically active cargoes into cells. Although concerted efforts have shed some light on the cellular internalization pathways of CPPs, quantification of CPP uptake has proved problematic. Here we describe an experimental approach that combines two powerful biophysical techniques, fluorescence activated cell sorting (FACS) and fluorescence correlation spectroscopy (FCS), to directly, accurately and precisely measure the cellular uptake of fluorescently-labeled molecules. This rapid and technically simple approach is highly versatile and can readily be applied to characterize all major CPP properties that normally require multiple assays, including amount taken up by cells (in moles/cell), uptake efficiency, internalization pathways, intracellular distribution, intracellular degradation and toxicity threshold. The FACS-FCS approach provides a means for quantifying any intracellular biochemical entity, whether expressed in the cell or introduced exogenously and transported across the plasma membrane. (C) 2016 The Authors. Published by Elsevier B.V.
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