Title
Population-based analysis of cell-penetrating peptide uptake using a microfluidic droplet trapping array
Document Type
Article
Publication Date
5-1-2019
Abstract
Cell-penetrating peptides (CPPs) have garnered significant attention as a method to introduce reporters and therapeutics into intact cells. While numerous studies have been performed identifying new CPP sequences, relatively little is known about their uptake efficiency at the single-cell level. Here, a droplet microfluidic trapping array was used to characterize CPP uptake across a population of single intact cells. The microfluidic device allowed for facile and rapid isolation and analysis of single-cell fluorescence in a 787-member overhead trapping array with > 99% droplet trapping efficiency. The permeability efficiencies of four different CPPs were studied and compared in HeLa cells. Population analysis was performed using linkage hierarchical cluster analysis by R programming to bin cells into subpopulations expressing very low to very high peptide uptake efficiencies. CPP uptake was observed to be heterogeneous across the population of cells with peptide concentration and sequence both playing important roles in the diversity of CPP uptake, the overall peptide uptake efficiency, and the intracellular homogeneity of peptide distribution. This microfluidic-based analytical approach finds application in personalized medicine and provides new insight in the heterogeneity of CPP uptake which has the potential to affect both biosensor and drug internalization in intact cells. Graphical abstract .
Publication Source (Journal or Book title)
Analytical and bioanalytical chemistry
First Page
2729
Last Page
2741
Recommended Citation
Safa, N., Vaithiyanathan, M., Sombolestani, S., Charles, S., & Melvin, A. T. (2019). Population-based analysis of cell-penetrating peptide uptake using a microfluidic droplet trapping array. Analytical and bioanalytical chemistry, 411 (12), 2729-2741. https://doi.org/10.1007/s00216-019-01713-5