PMID- 19110103
OWN - NLM
STAT- MEDLINE
DCOM- 20090210
LR  - 20171116
IS  - 1873-4324 (Electronic)
IS  - 0003-2670 (Linking)
VI  - 632
IP  - 2
DP  - 2009 Jan 26
TI  - A highly sensitive enzyme immunoassay for evaluation of 2'-deoxycytidine plasma 
      level as a prognostic marker for breast cancer chemotherapy.
PG  - 266-71
LID - 10.1016/j.aca.2008.11.014 [doi]
AB  - A highly sensitive competitive enzyme immunoassay (EIA) has been developed and 
      validated for the determination of the plasma level of 2'-deoxycytidine (dCyd), 
      the potential prognostic marker for breast cancer chemotherapy. This assay 
      employed a monoclonal antibody that recognizes dCyd with a high specificity, and 
      5'-succinyl-dCyd (5'sdCyd) conjugate of bovine serum albumin (5'sdCyd-BSA) 
      immobilized onto microplate wells as a solid phase. The assay involved a 
      competitive binding reaction between dCyd, in plasma sample, and the immobilized 
      5'sdCyd-BSA for the binding sites of the anti-dCyd antibody. The bound antibody 
      was quantified with horseradish peroxidase-labeled anti-immunoglobulin second 
      antibody and 3,3',5,5'-tetramethylbenzidine as a peroxidase substrate. The 
      concentration of dCyd in the sample was quantified by its ability to inhibit the 
      binding of the antibody to the immobilized 5'sdCyd-BSA and subsequently the color 
      formation in the assay. The assay limit of detection was 8 nM and the effective 
      working range at relative standard deviations (R.S.D.s) of <or=10% was 20-800 nM. 
      No cross-reactivity from the structurally related nucleobases, nucleosides, and 
      nucleotides was observed in the proposed assay. Mean analytical recovery of added 
      dCyd was 98-100+/-3.2-8.2%. The precision of the assay was satisfactory; R.S.D. 
      was 3.4-4.2 and 4.3-8.9% for intra- and inter-assay precision, respectively. The 
      proposed EIA was compared favorably with HPLC method in its ability to accurately 
      measure dCyd spiked into plasma samples. The analytical procedure is convenient, 
      and one can analyze 200 samples per working day, facilitating the processing of 
      large-number batch of samples. The proposed EIA is expected to contribute in 
      further evaluation of dCyd as a prognostic marker for breast cancer chemotherapy 
      and elucidation of the role of dCyd in various biological and biochemical 
      systems.
FAU - Darwish, Ibrahim A
AU  - Darwish IA
AD  - Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud 
      University, Riyadh, Saudi Arabia. idarwish@ksu.edu.sa
FAU - Mahmoud, Ashraf M
AU  - Mahmoud AM
FAU - Aboul-Fadl, Tarek
AU  - Aboul-Fadl T
FAU - Al-Majed, Abdul-Rahman A
AU  - Al-Majed AR
FAU - Khalil, Nasr Y
AU  - Khalil NY
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20081117
PL  - Netherlands
TA  - Anal Chim Acta
JT  - Analytica chimica acta
JID - 0370534
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Biomarkers)
RN  - 0W860991D6 (Deoxycytidine)
RN  - 27432CM55Q (Serum Albumin, Bovine)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal/immunology
MH  - Biomarkers/*blood/metabolism
MH  - Blood Chemical Analysis/*methods
MH  - Breast Neoplasms/blood/*diagnosis/*drug therapy
MH  - Cattle
MH  - Cell Line, Tumor
MH  - Chromatography, High Pressure Liquid
MH  - Deoxycytidine/*blood/immunology/metabolism
MH  - Immunoenzyme Techniques/*methods
MH  - Mice
MH  - Prognosis
MH  - Rats
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - Serum Albumin, Bovine/metabolism
EDAT- 2008/12/27 09:00
MHDA- 2009/02/12 09:00
CRDT- 2008/12/27 09:00
PHST- 2008/08/15 00:00 [received]
PHST- 2008/11/04 00:00 [revised]
PHST- 2008/11/05 00:00 [accepted]
PHST- 2008/12/27 09:00 [entrez]
PHST- 2008/12/27 09:00 [pubmed]
PHST- 2009/02/12 09:00 [medline]
AID - S0003-2670(08)01920-X [pii]
AID - 10.1016/j.aca.2008.11.014 [doi]
PST - ppublish
SO  - Anal Chim Acta. 2009 Jan 26;632(2):266-71. doi: 10.1016/j.aca.2008.11.014. Epub 
      2008 Nov 17.