Synthesis and cellular uptake of cell delivering PNA-peptide conjugates

Chemical Communications

Volumn , Issue 26, 2005, Pages 3316-3318

  Díaz Mochón, Juan J ^a   Bialy, Laurent ^a   Watson, Jon ^a   Sánchez Martín, Rosario M ^a   Bradley, Mark ^a  

^a UNIVERSITY OF EDINBURGH   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

ARGININE; FLUORESCENT DYE; MONOMER; PEPTIDE; PEPTIDE NUCLEIC ACID;

ARTICLE; BINDING AFFINITY; CELL MEMBRANE PERMEABILITY; CELL TRANSPORT; CHEMICAL ANALYSIS; CONTROLLED STUDY; HUMAN; HUMAN CELL; PROTEIN BINDING; PROTEIN INTERACTION; PROTEIN STRUCTURE; STRUCTURE ANALYSIS; SYNTHESIS;

EID: 22544432253     PISSN: 13597345     EISSN: None     Source Type: Journal    
DOI: 10.1039/b503777h     Document Type: Article

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19. +.
20. 2O for 5 min, then increasing from 0% to 60% MeCN over 10 min.
21. 4 cells per well and incubated overnight. Then cells were washed with warm PBS buffer and preincubated in 350 μl serum free medium (SFM) at 37 °C for 30 min. Compounds 9-12 were mixed with serum free medium at a final concentration of 500 nM. To each well different samples of 9-12 were added and incubated at 37 °C for 2 h. Each experiment was performed in triplicate. The internalisation of free fluorescein, under the same conditions, was tested simultaneously and untreated cells were used as negative control. After incubation, cells were washed twice with PBS, harvested with trypsin/EDTA, washed again and resuspended in 1% FCS in PBS buffer. To analyze the internalisation of fluorescein-labeled PNA conjugates, cell-associated fluorescence was determined by flow cytometry analysis using a FACSAria flow cytometer (Becton Dickinson). A total of 10 000 events per sample were analyzed. FITC (530/30 nm) band pass filters were used for fluorescence analysis of the cell suspensions. None of the conjugates 9-12 assayed was found to be toxic as verified by an MTT toxicity and a trypan blue assay to determine cell viability (see Supplementary Information).