Intracellular delivery of universal proteins using a lysine headgroup containing cationic liposomes: deciphering the uptake mechanism.
An amino acid-based cationic lipid having a TFA counterion (trifluoroacetic acid counterion) in the lysine headgroup was used to deliver functional proteins into human cervical cancer cells, HeLa, in the presence of serum. Proteins used in the study were fluorescein isothiocyanate (FITC) labeled bovine serum albumin, mouse anti-F actin antibody [NH3], and goat anti mouse IgG conjugated with FITC. The formation of liposome/protein complexes was confirmed using native polyacrylamide gel electrophoresis. Furthermore, the complexes were characterized in terms of their size and zeta potential at different pH values and found to be responsive to changes in pH. The highest delivery efficiency of the liposome/albumin complexes was 99% at 37 °C. The liposomes effectively delivered albumin and antibodies as confirmed by confocal laser scanning microscopy (CLSM). Inhibition studies showed that the cellular uptake mechanism of the complexes was via caveolae-mediated endocytosis, and the proteins were subsequently released from either the early endosomes or the caveosomes as suggested by CLSM. Thus, lysine-based cationic liposomes can be a useful tool for intracellular protein delivery.
Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University (TWIns) , Shinjuku-ku, Tokyo 162-8480, Japan.
Satya Ranjan
SR
Sarker
Ryosuke
R
Hokama
Shinji
S
Takeoka
D000818:Animals; D002412:Cations / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D016503:Drug Delivery Systems; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D008239:Lysine / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D012710:Serum Albumin, Bovine / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism
D002412:Cations; D008081:Liposomes; D012710:Serum Albumin, Bovine; D008239:Lysine