Protein transduction by lipidic peptide dendrimers.
We investigated the potential of a new family of lipidic peptide dendrimers in protein transduction into cultured cells. Dendrimer-protein interaction was determined by gel retardation assays using purified recombinant protein. To assess intracellular protein delivery, two marker proteins were used: recombinant firefly luciferase and a Cy3-labeled monoclonal antibody to the c-myc proto-oncogene. Protein delivery was determined by luciferase assays and fluorescence microscopy, respectively. While there was minimal delivery of luciferase or antibody in the absence of the dendrimers, the latter increased protein delivery substantially. Luciferase delivery was concentration and cell type-dependent; the efficiency of delivery also varied with the number of terminal amino groups on the dendrimers. In previous reports, we showed that these dendrimers could be used for gene and drug delivery; the data we report herein suggest that they may also be capable of intracellular protein delivery. This finding has important implications for the use of these dendrimers in protein therapeutics and vaccinology.
Bayele
Henry K
HK
Department of Biochemistry and Molecular Biology, University College London, London NW3 2PF, United Kingdom. h.bayele@medsch.ucl.ac.uk
Ramaswamy
Chandrasekaran
C
Wilderspin
Andrew F
AF
Srai
Kaila S
KS
Toth
Istvan
I
Florence
Alexander T
AT
D000911:Antibodies, Monoclonal / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D050091:Dendrimers / Q000138:chemical synthesis / Q000737:chemistry* / Q000302:isolation & purification / Q000633:toxicity; D004337:Drug Carriers / Q000737:chemistry*; D016503:Drug Delivery Systems; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay; D005456:Fluorescent Dyes; D005982:Glutathione Transferase / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D017785:Photons; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000090063:Proto-Oncogene Mas; D011994:Recombinant Proteins / Q000378:metabolism; D013057:Spectrum Analysis / Q000379:methods*; D014966:Xanthenes
D000911:Antibodies, Monoclonal; D050091:Dendrimers; D004337:Drug Carriers; D005456:Fluorescent Dyes; C000715470:MAS1 protein, human; D011506:Proteins; D000090063:Proto-Oncogene Mas; D011994:Recombinant Proteins; D014966:Xanthenes; C034657:Texas red; D008156:Luciferases; D005982:Glutathione Transferase