Prostate cancer is the most common malignant tumor in men, whose incidence significantly differs geograph-ically and increases with age. For rapid diagnostics, new tumor markers with higher prognostic relevance are still being sought. A very promising candidate molecule is the amino acid sarcosine. The aim of this work was to design a highly sensitive photometric detection system for the sarcosine determination. An original, yet unpublished, methodology for determining the amino acid sarcosine using Trinder's reaction has been proposed. Absorbance dependence on sarcosine concentration at 546 nm is linear over a total range of 0–1000 μM (r≥ 0.99) with limit of detection (LOD) of 6 μM and limit of quantification (LOQ) of 20 μM. The suggested procedure allows to ana-lyze the content of sarcosine in real urine specimens at micromolar concentrations.
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Presence of mutated and/or structurally modified (e.g., denatured, aggregated) protein p53 form is associated with several disorders such as Alzheimer's disease, Parkinson's disease, prion diseases, and many types of tumours. The aim of this work was to distinguish native, denatured and aggregated form of full-length p53 by flow injection analysis coupled with electrochemical detector (FIA-ED). Firstly FIA-ED method used for protein native form determination was optimized (detection limit 45.8 amol per 5 mul injection; 3 x S/N). In addition the technique was applied to identify p53 structural forms (denatured and aggregated). It was found out that denatured form provides about three times higher electrochemical response (protein structure unfolding, approach of more electroactive centers - aminoacid residues - towards electrode surface) in comparison with native form. On the other hand, aggregated form offers lower response (steric eclipse of electroactive protein parts) when compared with the signal of native form. The obtained data show that we are not only able to sensitively determine native, denatured, and aggregated structural forms of p53 protein but also to distinguish them.
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