Development and characterization of a monoclonal antibody with cross-reactivity towards uracil and thymine, and its potential use in screening patients treated with 5-fluorouracil for possible risks.
BACKGROUND
Dihydropyrimidine dehydrogenase (DPD) is the initial and rate-limiting enzyme in catabolism of pyrimidines including 5-fluorouracil. There have been efforts to isolate a monoclonal antibody that will bind selectively to pyrimidine and can be used to measure the concentration of pyrimidine in blood and/or in urine that may reflect the activity of dihydropyrimidine dehydrogenase. However, the monoclonal antibodies selective to pyrimidine have not been available.


METHODS
Using 1-carboxymethyl-uracil as a hapten, in which steric conformation of uracil is thought to be well maintained, extensive screening was done to isolate a monoclonal antibody specific to uracil.


RESULTS
We established the first monoclonal antibody that reacted with uracil and thymine but not with pseudouridine, dihydrouracil, dihydrothymine, cytosine, uridine, or N-carbamyl-beta-alanine at the concentration of 100 microg/ml.


CONCLUSIONS
The monoclonal antibody can be used to develop a simple screening assay for patients with dihydropyrimidine dehydrogenase deficiency. This may increase the safety of 5-fluorouracil treatment.
Honda
Teruko
T
Institute for Health Sciences, Tokushima Bunri University, Nishihama, Yamashiro-cho, Tokushima 770-8514, Japan.
Inagawa
Hiroyuki
H
Fukushima
Masakazu
M
Moriyama
Akira
A
Soma
Gen-Ichiro
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000075202:Contraindications; D003429:Cross Reactions / Q000276:immunology; D042943:Dihydrouracil Dehydrogenase (NADP); D004791:Enzyme Inhibitors / Q000276:immunology; D005472:Fluorouracil / Q000009:adverse effects*; D006801:Humans; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D010088:Oxidoreductases / Q000172:deficiency* / Q000378:metabolism; D011743:Pyrimidines / Q000276:immunology; D012307:Risk Factors; D013941:Thymine / Q000276:immunology* / Q000652:urine; D014498:Uracil / Q000276:immunology* / Q000652:urine
D000911:Antibodies, Monoclonal; D004791:Enzyme Inhibitors; D011743:Pyrimidines; D014498:Uracil; D010088:Oxidoreductases; D042943:Dihydrouracil Dehydrogenase (NADP); C030986:pyrimidine; D013941:Thymine; D005472:Fluorouracil