2 Materials
2.1 Preparation
of Lipid-Based Vaccine
Carriers
1. Trimethylpolyprenylammonium iodides with chain lengths
from 6 to 8 or 10 to 14 isoprene units—PTAI-6-8 or PTAI10-14, respectively or trimethylpolyprenylammonium iodides
with chain lengths of 7 or 11 isoprene units—PTAI-7 or PTAI11 (see Note 1). PTAI have been synthesized as described
earlier [15]. PTAI and their use are subject of patents
no. 230096 (System of nucleic acid carriers, method for preparation of the system and their applications), 211824 (Trimethylamine derivatives of poly-cis and poly-trans line isoprene oligomers,
method for their production and use), 231158 (Vaccine, pharmaceutical composition, carrier of nucleic acids and other biologically active substances, application of the composition in
production of the vaccine and application of cationic derivatives
Fig. 2 Structure and efficiency of trimethylpolyprenylammonium iodides (PTAI)
as lipofecting mixtures components. (a) Structure of PTAI, n corresponds to the
number of isoprenoid units in the polyprenyl chain; in this protocol n ¼ 6–8 or
10–14 or 7 or 11. (b, c) Efficiency of plasmid DNA (pEGFP-C1 encoding EGFP
enhanced green fluorescent protein) transfer into DU145 cells in the absence (b)
and in the presence of serum (c) in DMEM F-12 Ham medium with PTAI11 + DOPE + DC-cholesterol (1:1:1 molar ratio) reagent. Phase contrast (left
panel) and epifluorescence microscopy with FITC filter (right panel) images.
Scale bar 100 μm
Lipofection-Based Delivery of DNA Vaccines
393
2.1 Preparation
of Lipid-Based Vaccine
Carriers
1. Trimethylpolyprenylammonium iodides with chain lengths
from 6 to 8 or 10 to 14 isoprene units—PTAI-6-8 or PTAI10-14, respectively or trimethylpolyprenylammonium iodides
with chain lengths of 7 or 11 isoprene units—PTAI-7 or PTAI11 (see Note 1). PTAI have been synthesized as described
earlier [15]. PTAI and their use are subject of patents
no. 230096 (System of nucleic acid carriers, method for preparation of the system and their applications), 211824 (Trimethylamine derivatives of poly-cis and poly-trans line isoprene oligomers,
method for their production and use), 231158 (Vaccine, pharmaceutical composition, carrier of nucleic acids and other biologically active substances, application of the composition in
production of the vaccine and application of cationic derivatives
Fig. 2 Structure and efficiency of trimethylpolyprenylammonium iodides (PTAI)
as lipofecting mixtures components. (a) Structure of PTAI, n corresponds to the
number of isoprenoid units in the polyprenyl chain; in this protocol n ¼ 6–8 or
10–14 or 7 or 11. (b, c) Efficiency of plasmid DNA (pEGFP-C1 encoding EGFP
enhanced green fluorescent protein) transfer into DU145 cells in the absence (b)
and in the presence of serum (c) in DMEM F-12 Ham medium with PTAI11 + DOPE + DC-cholesterol (1:1:1 molar ratio) reagent. Phase contrast (left
panel) and epifluorescence microscopy with FITC filter (right panel) images.
Scale bar 100 μm
Lipofection-Based Delivery of DNA Vaccines
393