Microfluidic tool based on the antibody-modified paramagnetic particles for detection of 8-hydroxy-2'-deoxyguanosine in urine of prostate cancer patients.
Guanosine derivatives are important for diagnosis of oxidative DNA damage including 8-hydroxy-2'-deoxyguanosine (8-OHdG) as one of the most abundant products of DNA oxidation. This compound is commonly determined in urine, which makes 8-OHdG a good non-invasive marker of oxidation stress. In this study, we optimized and tested the isolation of 8-OHdG from biological matrix by using paramagnetic particles with an antibody-modified surface. 8-OHdG was determined using 1-naphthol generated by alkaline phosphatase conjugated with the secondary antibody. 1-Naphthol was determined by stopped flow injection analysis (SFIA) with electrochemical detector using a glassy carbon working electrode and by stationary electrochemical detection using linear sweep voltammetry. A special modular electrochemical SFIA system which needs only 10  μL of sample including working buffer for one analysis was completely designed and successfully verified. The recoveries in different matrices and analyte concentration were estimated. Detection limit (3 S/N) was estimated as 5  pg/mL of 8-OHdG. This method promises to be very easily modified to microfluidic systems as "lab on valve". The optimized method had sufficient selectivity and thus could be used for determination of 8-OHDG in human urine and therefore for estimation of oxidative DNA damage as a result of oxidation stress in prostate cancer patients.
Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Brno, Czech Republic.
Ondrej
O
Zitka
Sona
S
Krizkova
Ludmila
L
Krejcova
David
D
Hynek
Jaromir
J
Gumulec
Michal
M
Masarik
Jiri
J
Sochor
Vojtech
V
Adam
Jaromir
J
Hubalek
Libuse
L
Trnkova
Rene
R
Kizek
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000368:Aged; D000469:Alkaline Phosphatase; D000906:Antibodies; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000652:urine; D055664:Electrochemical Techniques / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D017022:Flow Injection Analysis; D006801:Humans; D059346:Magnets; D008297:Male; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008863:Microspheres; D008875:Middle Aged; D009284:Naphthols; D018384:Oxidative Stress; D011471:Prostatic Neoplasms / Q000652:urine*; D012371:Robotics / Q000295:instrumentation / Q000379:methods
D000906:Antibodies; D009284:Naphthols; C029350:1-naphthol; D000080242:8-Hydroxy-2'-Deoxyguanosine; D000469:Alkaline Phosphatase; D003849:Deoxyguanosine