The determination of urinary 3-methyladenine by immunoaffinity chromatography-monoclonal antibody-based ELISA: use in human biomonitoring studies.
A mouse monoclonal antibody (Mab) was prepared which showed high specificity for a potential marker of exposure to methylating carcinogens such as 3-methyladenine (3-MeAde). In a low-temperature (4 degrees C) ELISA a linear calibration curve was obtained between 3 and 50 fmol/well. In combination with an immunoaffinity (IA) column prepared from a 3-MeAde rabbit antiserum, the ELISA was used to determine 3-MeAde in urine. The IA-ELISA method was validated by comparison with results obtained by an IA-GC-MS method. The effect of consuming a low 3-MeAde diet on urinary 3-MeAde excretion was investigated in a human volunteer. Urine collected during a 'normal' diet exhibited the characteristic variation in 3-MeAde levels previously observed (9.5 micrograms/24 h, SD = 4.4, n = 5). In contrast, 3-MeAde excretion was markedly lower and less variable on days when the diet was closely controlled (0.63 microgram/24 h, SD = 0.08, n = 3). Dietary intake of 3-MeAde on the latter days was between 0.37 and 0.43 microgram/day, indicating that most, if not all, of the 3-MeAde seen in previous experiments was derived from the diet. The origin of dietary 3-MeAde is not known, but may be related to fumigant use. Dietary manipulation affords the possibility of carrying out model studies, in volunteers, on 3-MeAde intake and formation in vivo.
Prevost
V
V
International Agency for Research on Cancer, Lyon, France.
Shuker
D E
DE
Bartsch
H
H
Pastorelli
R
R
Stillwell
W G
WG
Trudel
L J
LJ
Tannenbaum
S R
SR
D000225:Adenine / Q000031:analogs & derivatives* / Q000652:urine; D000818:Animals; D000911:Antibodies, Monoclonal; D002273:Carcinogens; D002846:Chromatography, Affinity / Q000379:methods; D003429:Cross Reactions; D004032:Diet; D004784:Environmental Monitoring; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology
D000911:Antibodies, Monoclonal; D002273:Carcinogens; C025946:3-methyladenine; D000225:Adenine