Abstract
Abstract Background and Aims To determine the concentration of Alpha-2-macroglobulin (α2M) and its activity in patients treated with peritoneal dialysis (PD) and healthy subjects (H). Method a cross-sectional study included 50 PD and 30 H, in which the concentration of active α2M molecules was determined. The measurement is based on covalent binding of α2M and trypsin and zymography with gelatin incorporated into the electrophoretic gel. Results The intensity of proteolytic bands seen with native α2M was analyzed in relation to the concentration of α2M in patients with PD with less and / or more than three peritonitis. Linear dependence was only visible in the case of the upper range, which originates from the intact α2M-trypsin complex. The highest coefficient of variation is calculated for the smallest α2M concentration (4.2%). The signal intensities originating from α2M (H) were higher compared to PD, with the PD group being lower in patients with higher peritonitis, and the difference was more pronounced as concentration increased, resulting in a lower slope of dependence. The proposed method can reliably measure the amount of α2M originating in a group H, while the quantification of α2M in PD groups that may have structurally altered α2M can be seen more for condition estimation than for accurate measurements. The zymographic test in this experiment allows us to investigate the relationship of the concentration, structure and function of α2M in different pathological conditions, as well as in the presence of modifying agents that can accompany them. Conclusion A simple method for the determination of α2M concentration and function by zymography has been developed, which can be used to examine residual α2M activity after partial denaturation or structural modification due to disease.
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