The dynamic light scattering (DLS) technique was applied in order to assess the
zeta potential of the plasma membrane of human cells. At pH 7.4, the cell zeta
potential for different types of cells showed variations over a wide range and
was equal to –19.4 ± 0.8 mV for HeLa cells and –31.8 ± 1.1 mV for
erythrocytes. The difference could presumably be attributed to the differences
in the biochemical composition of the cell plasma membrane. As a result of the
heating of HeLa cells, the zeta potential shifted towards more negative voltages
by 4.2 mV. An increase in the zeta potential correlated with an increase in the
content of phosphatidylserine on the cell surface, which is considered to be an
early marker of apoptosis. The DLS technique was also used to study the
interactions between the cells and membranotropic polymers, such as polycations
and nonionogenic Pluronic L121.
We designed an electrochemical sensor based on a carbon nanotube modified electrode (ME) to analyze DNA-cleaving activity. The cleavage of high molecular weight DNA resulted in an increase in the oxidation current from DNA guanine nucleotides due to a change in DNA adsorptive behavior on the surface of the ME. DNA digestion with DNAse I was accompanied by a linear increase in the DNA signal in proportion to the enzyme activity. We then proposed an assay based on the sensor for the direct assessment of the total deoxyribonuclease activity of blood serum as well as the separate detection of DNAse I and DNA abzymes. The assay was applied to analyze deoxyribonucleases in sera from 21 healthy donors and 17 patients with autoimmune thyroiditis. Our results show that the response of the sensor to DNA cleavage by blood deoxyribonucleases is a promising diagnostic criterion for autoimmune thyroiditis. This sensor can be implemented in a disposable screen-printed electrode format for application in clinical laboratories.
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