Premium
Microfluidic tool based on the antibody‐modified paramagnetic particles for detection of 8‐hydroxy‐2′‐deoxyguanosine in urine of prostate cancer patients
Author(s) -
Zitka Ondrej,
Krizkova Sona,
Krejcova Ludmila,
Hynek David,
Gumulec Jaromir,
Masarik Michal,
Sochor Jiri,
Adam Vojtech,
Hubalek Jaromir,
Trnkova Libuse,
Kizek Rene
Publication year - 2011
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201100430
Subject(s) - detection limit , dna oxidation , 8 hydroxy 2' deoxyguanosine , chemistry , deoxyguanosine , chromatography , analyte , urine , linear sweep voltammetry , cyclic voltammetry , electrochemistry , oxidative stress , dna , dna damage , electrode , biochemistry
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.