title: Aberrant sialylation of a prostate-specific antigen: Electrochemical label-free glycoprofiling in prostate cancer serum samples.
abstract: Electrochemical detection method allowing to detect prostate-specific antigen (PSA), a biomarker of prostate cancer (PCa), with PSA glycoprofiling was applied in an analysis of PCa serum samples for the first time. Electrochemical impedance spectroscopy (EIS) as a label-free method with immobilized anti-PSA was applied for PSA detection and lectins to glycoprofile captured PSA on the same surface. A proper choice of blocking agent providing high selectivity of biosensor detection with the immobilized anti-PSA antibody was done. The biosensor could detect PSA down to 100 ag/mL with a linear concentration working range from 100 ag/mL up to 1 μg/mL, i.e. 10 orders of concentration magnitude and the sensitivity of (5.5 ± 0.2)%/decade. The results showed that a commercial carbo-free blocking solution was the best one, reducing non-specific binding 55-fold when compared to the immunosensor surface without any blocking agent applied, while allowing to detect PSA. The biosensor response obtained after addition of lectin (i.e. proportional to the amount of a particular glycan on PSA) divided by the biosensor response obtained after incubation with a sample (i.e. proportional to the PSA level in the sample) was applied to distinguish serum samples of PCa patients from those of healthy individuals. The results showed that Maackia amurensis agglutinin (MAA) recognizing α-2,3-terminal sialic acid can be applied to distinguish between these two sets of samples since the MAA/PSA response obtained from the analysis of the PCa samples was significantly higher (5.3-fold) compared to the MAA/PSA response obtained by the analysis of samples from healthy individuals. Thus, combined analysis of serological PSA levels together with PSA glycoprofiling of aberrant glycosylation of PSA (i.e. increase in the level of α-2,3-terminal sialic acid) has a potential to improve detection of PCa.
authors: Department of Glycobiotechnology, Institute of Chemistry, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava, 845 38, Slovak Republic.
authors: Dominika
authors: D
authors: Pihikova
authors: Centre for Advanced Materials, Qatar University, P.O. Box 2713, Doha, Qatar.
authors: Peter
authors: P
authors: Kasak
authors: Private Urological Ambulance, Piaristicka 6, Trencin, 911 01, Slovak Republic.
authors: Petra
authors: P
authors: Kubanikova
authors: Private Urological Ambulance, Piaristicka 6, Trencin, 911 01, Slovak Republic.
authors: Roman
authors: R
authors: Sokol
authors: Department of Glycobiotechnology, Institute of Chemistry, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava, 845 38, Slovak Republic. Electronic address: Jan.Tkac@savba.sk.
authors: Jan
authors: J
authors: Tkac
mesh_terms: D015374:Biosensing Techniques; D058266:Dielectric Spectroscopy; D055664:Electrochemical Techniques; D006031:Glycosylation; D006801:Humans; D008297:Male; D019158:N-Acetylneuraminic Acid / Q000097:blood*; D017430:Prostate-Specific Antigen / Q000097:blood*; D011471:Prostatic Neoplasms / Q000097:blood*
chemical_list: D017430:Prostate-Specific Antigen; D019158:N-Acetylneuraminic Acid
keywords: Biosensors; Electrochemical impedance spectroscopy; Lectins; Prostate cancer; Prostate-specific antigen