Voltammetric analysis of glycoproteins containing sialylated and neutral glycans at pyrolytic graphite electrode.

Recently, it was described that neutral glycans can be distinguished from those containing sialic acid at the mercury electrode after modification with osmium(VI) N,N,N',N'-tetramethylethylenediamine (Os(VI)tem). Our work shows the possibility of studying glycans and glycoproteins at pyrolytic graphite electrodes depending on thepresence of sialic acid. Short glycans, glycans released from glycoproteins, and glycoproteins themselves yielded similar voltammetric responses after their modification by Os(VI)tem.

Influence of Protein Modification and Glycosylation in the Catalytic Hydrogen Evolution Reaction of Avidin and Neutravidin: An Electrochemical Analysis.

To investigate glycans' influence on the behavior of glycoproteins on charged surfaces, avidin and its nonglycosylated and neutralized version neutravidin were studied by label-free chronopotentiometric stripping (CPS) analysis and alternating current voltammetry combined with a mercury electrode. Despite neutravidin's and avidin's similar size and structure, their CPS responses differed due to the different amounts of catalytically active free amino groups of lysine and arginine residues. Acetylation of the proteins resulted in the suppression of their CPS responses by almost four times for avidin and by about 50 % for neutravidin, respectively.

Cyclic and square wave voltammetry of chitooligosaccharides modified by osmium(VI) tetramethylethylenediamine.

Compounds containing vicinal diol (glycol) groups, including saccharides, could be modified with sixvalent osmium complexes with nitrogenous ligands, particularly with N,N,N',N'-tetramethylethylenediamine (Os(VI)tem). The modification products are electrochemically active. Here we show that aminosaccharides can also be modified by Os(VI)tem.

Vicinal Diol-Tethered Nucleobases as Targets for DNA Redox Labeling with Osmate Complexes.

Six-valent osmium (osmate) complexes with nitrogenous ligands have previously been used for the modification and redox labeling of biomolecules involving vicinal diol moieties (typically, saccharides or RNA). In this work, aliphatic (3,4-dihydroxybutyl and 3,4-dihydroxybut-1-ynyl) or cyclic (6-oxo-6-(cis-3,4-dihydroxypyrrolidin-1-yl)hex-2-yn-1-yl, PDI) vicinal diols are attached to nucleobases to functionalize DNA for subsequent redox labeling with osmium(VI) complexes. The diol-linked 2'-deoxyribonucleoside triphosphates were used for the polymerase synthesis of diol-linked DNA, which, upon treatment with K OsO and bidentate nitrogen ligands, gave the desired Os-labeled DNA, which were characterized by means of the gel-shift assay and ESI-MS.

Distinguishing the glycan isomers 2,3-sialyllactose and 2,6-sialyllactose by voltammetry after modification with osmium(VI) complexes.

Altered glycosylation is a universal feature of cancer cells and certain glycans are well-known markers of tumor progression. In this work we studied two glycan isomers, 2,3-sialyllactose (3-SL) and 2,6-sialyllactose (6-SL), frequently appearing in glycoproteins connected with cancer. A combination of square wave voltammetry and glycan modification with osmium(VI) N,N,N',N'-tetramethylethylenediamine (Os(VI)tem) allowed to distinguish between these regioisomers, since the 6-SL molecule can bind three Os(VI), while the 3-SL only two Os(VI) moieties, as experiments using capillary electrophoresis, inductively coupled plasma mass spectrometry and thin layer chromatography showed.

Immunoassays of chemically modified polysaccharides, glycans in glycoproteins and ribose in nucleic acids.

Glycosylation of proteins plays an important role in health and diseases. At present new simple and inexpensive methods of glycoprotein analysis are sought. We developed a monoclonal antibody Manost 2.

Voltammetric determination of Os(VI)-modified oligosaccharides at nanomolar level.

Glycoproteins participate in various biological events, including disease progression. Currently, there is a pressing need for development of new simple and inexpensive methods for glycoprotein carbohydrate component (mostly oligosaccharides, OLSs) analysis and electrochemical methods were little applied in their analysis. Polysaccharides and OLS were long time considered as electroinactive compounds.

Electrocatalysis in proteins, nucleic acids and carbohydrates.

The ability of proteins to catalyze hydrogen evolution has been known for more than 80 years, but the poorly developed d.c. polarographic "pre-sodium wave" was of little analytical use.

Electrocatalytic detection of polysaccharides at picomolar concentrations.

Electroinactive polysaccharides (PS) modified by osmium(VI) complexes with nitrogenous ligands produce redox couples at carbon and mercury electrodes. We show that PS adducts with Os(VI) 2,2'-bipyridine produce at ~-1.2 V (against Ag/AgCl/3 M KCl electrode) an additional peak at mercury and solid amalgam electrodes.

"Multicolor" electrochemical labeling of DNA hybridization probes with osmium tetroxide complexes.

Labeling of oligonucleotide reporter probes (RP) with electroactive markers has frequently been utilized in electrochemical detection of DNA hybridization. Osmium tetroxide complexes with tertiary amines (Os,L) bind covalently to pyrimidine (predominantly thymine) bases in DNA, forming stable, electrochemically active adducts. We propose a technique of electrochemical "multicolor" DNA coding based on RP labeling with Os,L markers involving different nitrogenous ligands (such as 2,2' -bipyridine, 1,10-phenanthroline derivatives or N,N,N',N'-tetramethylethylenediamine).