HIGH SELECTIVITY OF POLYCLONAL ANTIBODIES AGAINST DNA MODIFIED BY DIASTEREOMERIC BENZO[C]PHENANTHRENE-3,4-DIOL-1,2-EPOXIDES

被引:5
作者
BUTCH, ER
LAU, HHS
SHAW, KL
SMOLAREK, TA
SCHMEROLD, I
ANDERSON, JN
BAIRD, WM
YAGI, H
JERINA, DM
机构
[1] PURDUE UNIV,SCH PHARM & PHARMACAL SCI,DEPT MED CHEM,W LAFAYETTE,IN 47907
[2] PURDUE UNIV,SCH SCI,DEPT BIOL SCI,W LAFAYETTE,IN 47907
[3] NIDDK,BIOORGAN CHEM LAB,BETHESDA,MD 20892
关键词
D O I
10.1093/carcin/13.5.895
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Polyclonal antibodies were developed in New Zealand White rabbits against DNA modified with diastereomeric benzo[c]-phenanthrene-3,4-diol-1,2-epoxide (B[c]PhDE)-1 (4-hydroxyl and epoxide cis) and B[c]PhDE-2 (4-hydroxyl and epoxide trans). Antiserum developed against B[c]PhDE-2-DNA was stereoselective. In competitive ELISA assays using wells coated with 160 fmol B[c]PhDE-2-DNA adducts, B[c]PhDE-2-DNA gave 50% inhibition at 200 fmol adducts/well. B[c]PhDE-1-DNA required a 10-fold higher amount of adducts/well to give 50% inhibition. Benzo[a]pyrene-7,8-diol-9,10-epoxide-2-DNA and 7,12-dimethylbenz[a]-anthracene-3,4-diol-1,2-epoxide-1-DNA caused only a 30% inhibition even at the highest doses tested (> 4000 fmol adducts/well). For antiserum developed against B[c]PhDE-1-DNA, 50% inhibition required 570 fmol B[c]PhDE-1-DNA adducts in wells coated with 100 fmol B[c]PhDE-1-DNA adducts. 7,12-Dimethylbenz[a]anthracene-3,4-diol-1,2-epoxide-1-DNA and B[c]PhDE-2-DNA were also effective competitors: they caused 50% inhibition at 1900 and 1800 fmol adducts/well respectively. In contrast, benzo[a]-pyrene-7,8-diol-9,10-epoxide-2-DNA gave no inhibition at the highest dose of competitor tested (4050 fmol adducts/well). Antisera from three rabbits immunized, with B[c]PhDE-2-DNA demonstrated similar antigen specificities. The properties of these antisera differ from those reported previously for antibodies developed against benzo[a]-pyrene-DNA in that they show selectivity for DNA modified by specific hydrocarbon diolepoxides, in one case for B[c]PhDE-2-DNA and in the other for B[c]PhDE-DNA or 7,12-dimethylbenz[a]anthracene-3,4-diol-1,2-epoxide-1-DNA. The specificity of these antisera will facilitate analysis of the modification of DNA by different polycyclic aromatic hydrocarbon diolepoxides.
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页码:895 / 899
页数:5
相关论文
共 25 条
[1]   CHEMICAL CHARACTERIZATION OF DNA ADDUCTS DERIVED FROM THE CONFIGURATIONALLY ISOMERIC BENZO[C]PHENANTHRENE-3,4-DIOL 1,2-EPOXIDES [J].
AGARWAL, SK ;
SAYER, JM ;
YEH, HJC ;
PANNELL, LK ;
HILTON, BD ;
PIGOTT, MA ;
DIPPLE, A ;
YAGI, H ;
JERINA, DM .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1987, 109 (08) :2497-2504
[2]  
BIGGER CAH, 1983, CANCER RES, V43, P5647
[3]  
DIPPLE A, 1983, CANCER RES, V43, P4132
[4]   OPTICALLY-ACTIVE BENZO[C]PHENANTHRENE DIOL EPOXIDES BIND EXTENSIVELY TO ADENINE IN DNA [J].
DIPPLE, A ;
PIGOTT, MA ;
AGARWAL, SK ;
YAGI, H ;
SAYER, JM ;
JERINA, DM .
NATURE, 1987, 327 (6122) :535-536
[5]  
HSU IC, 1981, CANCER RES, V41, P1091
[6]  
Jerina D M, 1986, Adv Exp Med Biol, V197, P11
[7]  
JERINA DM, 1976, DRUG METABOLISM MICR, P13
[8]  
KOREEDA M, 1978, SCIENCE, V199, P778, DOI 10.1126/science.622566
[9]   DETECTION AND IDENTIFICATION OF BENZO[A]PYRENE DNA ADDUCTS BY [S-35] PHOSPHOROTHIOATE LABELING AND HPLC [J].
LAU, HHS ;
BAIRD, WM .
CARCINOGENESIS, 1991, 12 (05) :885-893
[10]   SYNTHESIS OF THE ACTIVE DIOL EPOXIDE METABOLITES OF THE POTENT CARCINOGENIC HYDROCARBON 7,12-DIMETHYLBENZ[A]ANTHRENE [J].
LEE, H ;
HARVEY, RG .
JOURNAL OF ORGANIC CHEMISTRY, 1986, 51 (18) :3502-3507