Monoclonal antibodies and rabbit antisera recognizing 4-aminobiphenyl—DNA adducts and application to immunoaffinity chromatography

Groopman, John D.; Skipper, Paul L.; Donahue, Paul R.; Trudel, Laura J.; Wildschutte, Michael; Kadlubar, Fred F.; Tannenbaum, Steven R.

doi:10.1093/carcin/13.6.917

Cited by 8 publications

(3 citation statements)

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“…GC/MS data indicated an average DNA adduct level of 4.72/108 in liver tissues of mice treated with 10 mg/kg. Thus, these data also suggest that the Two monoclonal antisera (3D6 and 2E11) against N-acetyl-4-aminobiphenyl-modified bovine y-globulin and a polyclonal antiserum (BH2) against 4-ABP-deoxyguanosine-keyhole limpet hemocyanin were developed previously (6,15). With 4-ABP-dG, 750% was at 2 and 65 pmol for antisera 3D6 and 2E11, respectively.…”

Section: Discussionsupporting

confidence: 59%

Quantitative Immunohistochemical Analysis of 4-Aminobiphenyl-DNA in Cultured Cells and Mice: Comparison to Gas Chromatography/Mass Spectroscopy Analysis

Al-Atrash¹,

Zhang²,

Lin³

et al. 1995

Chem. Res. Toxicol.

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Two monoclonal antisera, 4C11 and 3C8, recognizing 4-aminobiphenyl (4-ABP)--DNA adducts were developed and characterized by competitive enzyme-linked immunosorbent assay (ELISA). Both antisera are highly specific for 4-ABP-DNA and, at the highest concentration tested, do not recognize the DNA adducts of several other aromatic amines tested including 1-aminopyrene, 8-nitro-1-aminopyrene, and 6-nitro-1-aminopyrene. An immunohistochemical method for detecting adducts was developed in R52 cells, a mouse NIH3T3 cell line expressing high levels of cytochrome P450 1A2. Quantitation of fluorescence labeling indicated a dose-related increase in staining in cells treated with 0, 6, 30, 60, and 300 microM 4-ABP. To apply the method to tissue samples, Balb/c mice were treated with 0, 4, 10, 20, 40, and 80 mg/kg 4-ABP and liver, bladder, and lung tissue analyzed by immunohistochemical staining of tissue sections. There was a dose-related increase in specific nuclear staining in liver and bladder tissues with no detectable staining in lung tissue. DNA from liver tissue was also analyzed by alkaline hydrolysis of 4-ABP, derivatization with pentafluoropropionic anhydride, and gas chromatography/mass spectroscopy analysis. A good correlation (r = 0.98, p < 0.0001) was found between DNA damage levels determined by the two methods. Based on adduct levels determined by GC/MS in both R52 cells and liver tissue, the immunohistochemical method has a limit of sensitivity of approximately 1 adduct/10(7-8) nucleotides. Immunohistochemistry should be useful for analysis of 4-ABP-DNA adducts in human tissue biopsies as well as exfoliated cells from the oral mucosa and urinary bladder.

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Section: Discussionsupporting

confidence: 59%

Quantitative Immunohistochemical Analysis of 4-Aminobiphenyl-DNA in Cultured Cells and Mice: Comparison to Gas Chromatography/Mass Spectroscopy Analysis

Al-Atrash¹,

Zhang²,

Lin³

et al. 1995

Chem. Res. Toxicol.

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“…Since the induction of PII enzymes is implicated in detoxification of environmental toxins like aflatoxin (Kensler et al, 1986; Roebuck et al, 1991; Groopman et al, 1992), the effects of a broccoli sprout infusion (BSI) were evaluated on urinary aflatoxin and phenanthrene metabolite excretion (Kensler et al, 2005). Study participants ( n = 200) were recruited from Qidong, People’s Republic of China, which has a high prevalence of hepatocellular carcinoma emanating from chronic hepatitis B infection and exposure to aflatoxins in the diet (Ross et al, 1992).…”

Section: Sulforaphane and Are-dependent Gene Inductionmentioning

confidence: 99%

Induction of Phase 2 Antioxidant Enzymes by Broccoli Sulforaphane: Perspectives in Maintaining the Antioxidant Activity of Vitamins A, C, and E

Boddupalli¹,

Mein²,

Lakkanna³

et al. 2012

Front. Gene.

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Consumption of fruits and vegetables is recognized as an important part of a healthy diet. Increased consumption of cruciferous vegetables in particular has been associated with a decreased risk of several degenerative and chronic diseases, including cardiovascular disease and certain cancers. Members of the cruciferous vegetable family, which includes broccoli, Brussels sprouts, cauliflower, and cabbage, accumulate significant concentrations of glucosinolates, which are metabolized in vivo to biologically active isothiocyanates (ITCs). The ITC sulforaphane, which is derived from glucoraphanin, has garnered particular interest as an indirect antioxidant due to its extraordinary ability to induce expression of several enzymes via the KEAP1/Nrf2/ARE pathway. Nrf2/ARE gene products are typically characterized as Phase II detoxification enzymes and/or antioxidant (AO) enzymes. Over the last decade, human clinical studies have begun to provide in vivo evidence of both Phase II and AO enzyme induction by SF. Many AO enzymes are redox cycling enzymes that maintain redox homeostasis and activity of free radical scavengers such as vitamins A, C, and E. In this review, we present the existing evidence for induction of PII and AO enzymes by SF, the interactions of SF-induced AO enzymes and proposed maintenance of the essential vitamins A, C, and E, and, finally, the current view of genotypic effects on ITC metabolism and AO enzyme induction and function.

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“…Several approaches, including immunoaffinity and column chromatography, as well as extraction with toluene or hexane were evaluated for removal of these impurities. Immunoaffinity columns containing immobilized ABPspecific monoclonal antibody were prepared as described (13). Reaction mixture containing 35S-TBM-[3H]dGuo-8-ABP was redissolved in THF (20 ,l4), methanol (100 p4), and H20 (10 ml), and aliquots were loaded onto immunoaffinity columns, the column was washed with 20 ml of H20, and then material was eluted with 20 ml of 100%o methanol.…”

Section: Methodsmentioning

confidence: 99%

Adduct detection by acylation with [35S]methionine:analysis of DNA adducts of 4-aminobiphenyl.

Sheabar

Morningstar

Wogan

1994

Proc. Natl. Acad. Sci. U.S.A.

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Reaction ofsyntheticN-(2'-deoxyguanosin-8-yl)-4-amlnobiphenyl Analysis of human DNA samples with existing methods can produce conflicting results (2, 10), emphasizing the utility of alternative methodologies. Our objective was to develop a method for detecting carcinogen-DNA adducts that could be broadly applicable to carcinogens representing diverse chemical classes, have sensitivity adequate to detect adduct levels resulting from ambient exposures, be capable of identifying individual adducts as well as mixtures resulting from complex exposures, have potential for quantifying adduct recovery and structural identification of specific carcinogen adducts, and utilize readily available reagents and equipment. The experimental strategy for a method addressing these objectives, based on chemical derivatization of mononucleosides, is summarized in Fig. 1. DNA is digested enzymatically and adducted nucleosides are separated from unmodified nucleosides either by reversed-phase chromatography, for bulk separation, or by immunoaffnmity purification of specific carcinogen adducts for which monoclonal antibodies are available. For quantification of adducts of specific carcinogens, adducted nucleosides isolated by immunoaffinity purification can be subjected directly to acylation with t-butoxycarbonyl-L-[35S]methionine, N-hydroxysuccinimidyl ester (35S-TBM-NHS), for identification and quantification.Alternatively, adducted nucleosides can be acylated as a mixture, after which total adduct levels can be determined by HPLC separation, and in parallel, specific carcinogen adducts can be determined by immunoaffinity purification with monoclonal antibodies. Nucleoside adducts are acylated with an 35S-TBM-NHS reagent previously used for protein labeling through reaction with primary amine groups (11,12). After the reaction under conditions optimized for each class of adducts, the derivatives are chromatographically separated by HPLC and radioactivity was quantified with a flow-through liquid scintillation detector. Yields at each step and overall recovery can be quantified through the use of internal standards and the acylation products identified by use of authentic reference nucleosides. MATERIALS AND METHODSChemicals were purchased as follows: 2'-deoxyguanosine and Sephadex LH-20 from Sigma; 4-aminobiphenyl, triethylamine (TEA), and N',N'-diisopropylcarbodiimide (DIC) from Aldrich; 2'-deoxy-1',2'-[3H]guanosine 5'-triphosphate

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Monoclonal antibodies and rabbit antisera recognizing 4-aminobiphenyl—DNA adducts and application to immunoaffinity chromatography

Cited by 8 publications

References 0 publications

Quantitative Immunohistochemical Analysis of 4-Aminobiphenyl-DNA in Cultured Cells and Mice: Comparison to Gas Chromatography/Mass Spectroscopy Analysis

Quantitative Immunohistochemical Analysis of 4-Aminobiphenyl-DNA in Cultured Cells and Mice: Comparison to Gas Chromatography/Mass Spectroscopy Analysis

Induction of Phase 2 Antioxidant Enzymes by Broccoli Sulforaphane: Perspectives in Maintaining the Antioxidant Activity of Vitamins A, C, and E

Adduct detection by acylation with [35S]methionine:analysis of DNA adducts of 4-aminobiphenyl.

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