Amyloid aggregation of proteins is usually associated with amyloid diseases. A distinct feature of protein aggregation is the increase of crossed β-sheet structures. Infrared attenuated-total-reflectance (IR-ATR) spectroscopy is a sensitive optical technique that has the potential to provide secondary structure characteristics of proteins even in complex biological samples. In this study we report the analysis of secondary structures of proteins, using the amide I band for the detection and quantification of amyloid fibrils in protein mixtures by use of IR-ATR techniques, at comparatively low sample concentrations. From the experimental results, an analytical model of the relationship between the IR spectra of protein mixtures and the individual mixture components was established using spectral-deconvolution procedures and curve-fitting methods. On the basis of this model, four ratios were shown to provide direct information on amyloid aggregated fibrils via the increase of crossed β-sheet structures in protein-mixture samples. In conclusion, this study confirms the utility of IR spectroscopy for analyzing protein mixtures and for identifying amyloid fibril information within such complex multi-component samples.

The synthesis of two types of viologen dendrimers with peripheral carboxyl groups is described. Their interaction with plasmid DNA and CT-DNA and the influence of time evolution and electrolyte on dendriplex formation have been electrochemically investigated. A negative potential shift appearing in the cyclic voltammograms of the dendrimers indicates dendriplex formation on the time scale of 15 to 19 minutes, i.e. similar to those determined empirically for other dendrimer types. The presence or absence of the negative potential shift can be used to check the stability towards sodium chloride and different cell growth media directing to sucrose for cell incubation experiments. The electrolyte content of commercially available cell growth media inhibits the dendriplex formation in solution prior to plasmid addition.Furthermore, a low salt stability of 20 mM sodium chloride for viologen dendriplexes has been confirmed, also recommending the use of lysosomotropic sucrose. The two types of viologen dendrimers have been combined with two plasmids differing in the number of base pairs. Four immortal cell lines have been tested to check the suitability of viologen dendriplexes as gene delivery systems. Probably due to the absence of terminal amino groups and endosomolytic substances only a small transfection efficiency of dendriplexes was achieved at low pH, generally excluding in vivo applications. With the larger pHSV-eGFP plasmid (5743 bp) no transfected cells were observed indicating a preference for shorter plasmids.