Self-assembled nanostructures
a self-assembled, nano-structure technology, applied in the direction of peptides, fusion polypeptides, chemistry apparatus and processes, etc., can solve the problems of unclear morphology, insufficient stability and biological activity of structure for nucleic acids delivery, etc., and achieve high purity
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example 1
Molecular Structure of Exemplary Self Assembling DNA Binding Peptides
[0183]The design described herein above led to the synthesis of the following PEGylated DNA binding peptides (FIG. 2).
Synthesis of Peptide Segment
[0184]The synthesis of the peptidic segment was achieved by conventional solid phase synthesis on Tentagel-R-RAM resin using the Fmoc protecting group strategy. The introduction of the spermine DNA binding unit at the C-terminus of the coiled-coil peptide was achieved by coupling the Boc protected 5-carboxyspermine to the 6-amine of a lysine residue. The PEG chain was introduced by direct coupling of a PEG2000 carboxylic acid to the free N-terminus of the peptide on solid phase to give 1, or by the conjugation of a commercial mPEG5000-maleimide to the coiled-coil peptide functionalize with a N-terminal cysteine residue to give 2. The individual monodisperse double strand NoLimits DNA fragments were purchased from Fermentas. The plasmid DNA templates were purchased from Ne...
example 2
Sedimentation Analysis
[0186]Equilibrium: The coiled-coil formation was then investigated further by analytical ultracentrifugation, which gives access to the absolute molecular weight of the aggregates in solution and subsequently their aggregation number. Equilibrium experiments were conducted at 23° C. and in 50 mM phosphate buffer containing 150 mM NaCl, pH 7.4. Three loading concentrations (18, 30 and 42 μM) were scanned at 230 nm, and sedimented to equilibrium at 20 krpm, 26.6 krpm, 33.3 krpm, and at 39.9 krpm, resulting in 12 scans which were globally fit to multiple models as described previously [Johnson et al., Biophys. J. 36(3): 575-588 (1981); Demeler et al., UltraScan A Comprehensive Data Analysis Software Package for Analytical Ultracentrifugation Experiments. Analytical Ultracentrifugation—Techniques and Methods. Royal Society of Chemistry: 2005]. The most appropriate model was chosen based on visual inspection of the residual run patterns and based on the best statist...
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