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Enhancing cellular uptake of GFP via unfolded supercharged protein tags
- Source :
- Biomaterials, Biomaterials, 34(17), 4360-4367. ELSEVIER SCI LTD
- Publication Year :
- 2013
-
Abstract
- One of the barriers to the development of protein therapeutics is effective delivery to mammalian cells. The proteins must maintain a careful balance of polar moieties to enable administration and distribution and hydrophobic character to minimize cell toxicity. Numerous strategies have been applied to this end, from appending additional cationic peptides to supercharging the protein itself, sometimes with limited success. Here we present a strategy that combines these methods, by equipping a protein with supercharged elastin-like polypeptide (ELP) tags. We monitored cellular uptake and cell viability for GFP reporter proteins outfitted with a range of ELP tags and demonstrated enhanced uptake that correlates with the number of positive charges, while maintaining remarkably low cytotoxicity and resistance to degradation in the cell. GFP uptake proceeded mainly through caveolae-mediated endocytosis and we observed GFP emission inside the cells over extended time (up to 48 h). Low toxicity combined with high molecular weights of the tag opens the way to simultaneously optimize cell uptake and pharmacokinetic parameters. Thus, cationic supercharged ELP tags show great potential to improve the therapeutic profile of protein drugs leading to more efficient and safer biotherapeutics. (C) 2013 Elsevier Ltd. All rights reserved.
- Subjects :
- MEDIATED DELIVERY
TUMOR-CELLS
Cell
Intracellular Space
Supercharged protein
02 engineering and technology
Protein tag
Green fluorescent protein
GENE DELIVERY
MAMMALIAN-CELLS
Cytotoxicity
ARGININE-RICH PEPTIDES
FUNCTIONAL PROTEINS
0303 health sciences
Microscopy, Confocal
Cell Death
021001 nanoscience & nanotechnology
FUSOGENIC PEPTIDE
Endocytosis
Cell biology
medicine.anatomical_structure
Biochemistry
Mechanics of Materials
Elastin-like polypeptide
0210 nano-technology
Fusion protein
Recombinant Fusion Proteins
Green Fluorescent Proteins
Molecular Sequence Data
Biophysics
Bioengineering
Gene delivery
Biology
GFP
Biomaterials
03 medical and health sciences
Cell Line, Tumor
Protein therapeutic
medicine
Humans
Viability assay
Amino Acid Sequence
CELLS IN-VITRO
030304 developmental biology
Protein Unfolding
Elastin
POLY-L-LYSINE
Ceramics and Composites
PENETRATING PEPTIDES
Peptides
Subjects
Details
- Language :
- English
- ISSN :
- 01429612
- Database :
- OpenAIRE
- Journal :
- Biomaterials 2013, 34: 4360-4367.
- Accession number :
- edsair.doi.dedup.....ad6c69ecaaaf7210c4ac81b4803ed5a4
- Full Text :
- https://doi.org/10.1016/j.biomaterials.2013.02.038