Measurement of benzo(a)pyrene-DNA adducts by enzyme immunoassays and radioimmunoassay

I. C. Hsu, M. C. Poirier, S. H. Yuspa, D. Grunberger, I. B. Weinstein, Robert H Yolken, C. C. Harris

Research output: Contribution to journalArticle

Abstract

Ultrasensitive enzymatic radioimmunoassay (USERIA) was compared to radioimmunoassay and enzyme-linked immunosorbent assay in determining the amount of benzo(a)pyrene [B(a)P] metabolite covalently bound to guanine in DNA. In the USERIA approach, DNA either with or without B(a)P metabolite modification was adsorbed on the wells of microtiter plates, and rabbit antiserum to B(a)P metabolite-modified DNA [B(a)P-DNA] was then added. Antibodies reacted specifically with the B(a)P-DNA attached to the surface of the plate. After reaction between goat anti-rabbit IgG conjugated to alkaline phosphatase and rabbit IgG bound to the solid phase, this specific antigen-antibody reaction was enzymatically amplified by alkaline phosphatase conversion of [ 3H]adenosine 5'monophosphate to [ 3H]adenosine. Following chromatographic separation from [ 3H]adenosine 5'-monophosphate, [ 3H]adenosine was measured by liquid scintillation counting. The amount of 3H]-adenosine formed was linearly related to the amount of B(a)P-DNA in 10 ng DNA attached to the solid phase. As little as 3 fmol of bound B(a)P metabolite can be detected by noncompetitive USERIA, while 10 fmol of the adducts in 25 μg DNA [1 B(a)P-DNA adduct per 7 X 10 6 nucleotides] can be measured by the competitive USERIA approach. Under our standard competitive procedure with 1 μg DNA in the antigen-antibody reaction mixture, USERIA is approximately 500-fold more sensitive than radioimmunoassay and 5-fold more sensitive than enzyme-linked immunosorbent assay for the detection of B(a)P-DNA adducts. These highly sensitive assays should be extremely useful for studies in DNA damage and repair by B(a)P metabolites as well as in studies on environmental exposure to this ubiquitous carcinogen.

Original languageEnglish (US)
Pages (from-to)1091-1095
Number of pages5
JournalCancer Research
Volume41
Issue number3
StatePublished - 1981
Externally publishedYes

Fingerprint

Benzo(a)pyrene
Immunoenzyme Techniques
Radioimmunoassay
DNA
Adenosine
Antigen-Antibody Reactions
Adenosine Monophosphate
Rabbits
Alkaline Phosphatase
Enzyme-Linked Immunosorbent Assay
benzo(a)pyrene-DNA adduct
Scintillation Counting
Environmental Exposure
Guanine
Goats
DNA Repair
Carcinogens
DNA Damage
Immune Sera
Nucleotides

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Hsu, I. C., Poirier, M. C., Yuspa, S. H., Grunberger, D., Weinstein, I. B., Yolken, R. H., & Harris, C. C. (1981). Measurement of benzo(a)pyrene-DNA adducts by enzyme immunoassays and radioimmunoassay. Cancer Research, 41(3), 1091-1095.

Measurement of benzo(a)pyrene-DNA adducts by enzyme immunoassays and radioimmunoassay. / Hsu, I. C.; Poirier, M. C.; Yuspa, S. H.; Grunberger, D.; Weinstein, I. B.; Yolken, Robert H; Harris, C. C.

In: Cancer Research, Vol. 41, No. 3, 1981, p. 1091-1095.

Research output: Contribution to journalArticle

Hsu, IC, Poirier, MC, Yuspa, SH, Grunberger, D, Weinstein, IB, Yolken, RH & Harris, CC 1981, 'Measurement of benzo(a)pyrene-DNA adducts by enzyme immunoassays and radioimmunoassay', Cancer Research, vol. 41, no. 3, pp. 1091-1095.
Hsu IC, Poirier MC, Yuspa SH, Grunberger D, Weinstein IB, Yolken RH et al. Measurement of benzo(a)pyrene-DNA adducts by enzyme immunoassays and radioimmunoassay. Cancer Research. 1981;41(3):1091-1095.
Hsu, I. C. ; Poirier, M. C. ; Yuspa, S. H. ; Grunberger, D. ; Weinstein, I. B. ; Yolken, Robert H ; Harris, C. C. / Measurement of benzo(a)pyrene-DNA adducts by enzyme immunoassays and radioimmunoassay. In: Cancer Research. 1981 ; Vol. 41, No. 3. pp. 1091-1095.
@article{d1b687b842c040d78e833808afda0859,
title = "Measurement of benzo(a)pyrene-DNA adducts by enzyme immunoassays and radioimmunoassay",
abstract = "Ultrasensitive enzymatic radioimmunoassay (USERIA) was compared to radioimmunoassay and enzyme-linked immunosorbent assay in determining the amount of benzo(a)pyrene [B(a)P] metabolite covalently bound to guanine in DNA. In the USERIA approach, DNA either with or without B(a)P metabolite modification was adsorbed on the wells of microtiter plates, and rabbit antiserum to B(a)P metabolite-modified DNA [B(a)P-DNA] was then added. Antibodies reacted specifically with the B(a)P-DNA attached to the surface of the plate. After reaction between goat anti-rabbit IgG conjugated to alkaline phosphatase and rabbit IgG bound to the solid phase, this specific antigen-antibody reaction was enzymatically amplified by alkaline phosphatase conversion of [ 3H]adenosine 5'monophosphate to [ 3H]adenosine. Following chromatographic separation from [ 3H]adenosine 5'-monophosphate, [ 3H]adenosine was measured by liquid scintillation counting. The amount of 3H]-adenosine formed was linearly related to the amount of B(a)P-DNA in 10 ng DNA attached to the solid phase. As little as 3 fmol of bound B(a)P metabolite can be detected by noncompetitive USERIA, while 10 fmol of the adducts in 25 μg DNA [1 B(a)P-DNA adduct per 7 X 10 6 nucleotides] can be measured by the competitive USERIA approach. Under our standard competitive procedure with 1 μg DNA in the antigen-antibody reaction mixture, USERIA is approximately 500-fold more sensitive than radioimmunoassay and 5-fold more sensitive than enzyme-linked immunosorbent assay for the detection of B(a)P-DNA adducts. These highly sensitive assays should be extremely useful for studies in DNA damage and repair by B(a)P metabolites as well as in studies on environmental exposure to this ubiquitous carcinogen.",
author = "Hsu, {I. C.} and Poirier, {M. C.} and Yuspa, {S. H.} and D. Grunberger and Weinstein, {I. B.} and Yolken, {Robert H} and Harris, {C. C.}",
year = "1981",
language = "English (US)",
volume = "41",
pages = "1091--1095",
journal = "Journal of Cancer Research",
issn = "0099-7013",
publisher = "American Association for Cancer Research Inc.",
number = "3",

}

TY - JOUR

T1 - Measurement of benzo(a)pyrene-DNA adducts by enzyme immunoassays and radioimmunoassay

AU - Hsu, I. C.

AU - Poirier, M. C.

AU - Yuspa, S. H.

AU - Grunberger, D.

AU - Weinstein, I. B.

AU - Yolken, Robert H

AU - Harris, C. C.

PY - 1981

Y1 - 1981

N2 - Ultrasensitive enzymatic radioimmunoassay (USERIA) was compared to radioimmunoassay and enzyme-linked immunosorbent assay in determining the amount of benzo(a)pyrene [B(a)P] metabolite covalently bound to guanine in DNA. In the USERIA approach, DNA either with or without B(a)P metabolite modification was adsorbed on the wells of microtiter plates, and rabbit antiserum to B(a)P metabolite-modified DNA [B(a)P-DNA] was then added. Antibodies reacted specifically with the B(a)P-DNA attached to the surface of the plate. After reaction between goat anti-rabbit IgG conjugated to alkaline phosphatase and rabbit IgG bound to the solid phase, this specific antigen-antibody reaction was enzymatically amplified by alkaline phosphatase conversion of [ 3H]adenosine 5'monophosphate to [ 3H]adenosine. Following chromatographic separation from [ 3H]adenosine 5'-monophosphate, [ 3H]adenosine was measured by liquid scintillation counting. The amount of 3H]-adenosine formed was linearly related to the amount of B(a)P-DNA in 10 ng DNA attached to the solid phase. As little as 3 fmol of bound B(a)P metabolite can be detected by noncompetitive USERIA, while 10 fmol of the adducts in 25 μg DNA [1 B(a)P-DNA adduct per 7 X 10 6 nucleotides] can be measured by the competitive USERIA approach. Under our standard competitive procedure with 1 μg DNA in the antigen-antibody reaction mixture, USERIA is approximately 500-fold more sensitive than radioimmunoassay and 5-fold more sensitive than enzyme-linked immunosorbent assay for the detection of B(a)P-DNA adducts. These highly sensitive assays should be extremely useful for studies in DNA damage and repair by B(a)P metabolites as well as in studies on environmental exposure to this ubiquitous carcinogen.

AB - Ultrasensitive enzymatic radioimmunoassay (USERIA) was compared to radioimmunoassay and enzyme-linked immunosorbent assay in determining the amount of benzo(a)pyrene [B(a)P] metabolite covalently bound to guanine in DNA. In the USERIA approach, DNA either with or without B(a)P metabolite modification was adsorbed on the wells of microtiter plates, and rabbit antiserum to B(a)P metabolite-modified DNA [B(a)P-DNA] was then added. Antibodies reacted specifically with the B(a)P-DNA attached to the surface of the plate. After reaction between goat anti-rabbit IgG conjugated to alkaline phosphatase and rabbit IgG bound to the solid phase, this specific antigen-antibody reaction was enzymatically amplified by alkaline phosphatase conversion of [ 3H]adenosine 5'monophosphate to [ 3H]adenosine. Following chromatographic separation from [ 3H]adenosine 5'-monophosphate, [ 3H]adenosine was measured by liquid scintillation counting. The amount of 3H]-adenosine formed was linearly related to the amount of B(a)P-DNA in 10 ng DNA attached to the solid phase. As little as 3 fmol of bound B(a)P metabolite can be detected by noncompetitive USERIA, while 10 fmol of the adducts in 25 μg DNA [1 B(a)P-DNA adduct per 7 X 10 6 nucleotides] can be measured by the competitive USERIA approach. Under our standard competitive procedure with 1 μg DNA in the antigen-antibody reaction mixture, USERIA is approximately 500-fold more sensitive than radioimmunoassay and 5-fold more sensitive than enzyme-linked immunosorbent assay for the detection of B(a)P-DNA adducts. These highly sensitive assays should be extremely useful for studies in DNA damage and repair by B(a)P metabolites as well as in studies on environmental exposure to this ubiquitous carcinogen.

UR - http://www.scopus.com/inward/record.url?scp=0019431506&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0019431506&partnerID=8YFLogxK

M3 - Article

C2 - 7459852

AN - SCOPUS:0019431506

VL - 41

SP - 1091

EP - 1095

JO - Journal of Cancer Research

JF - Journal of Cancer Research

SN - 0099-7013

IS - 3

ER -