BIOMAT Database Logo BIOMAT Database Logo

Interlaboratory comparison of antisera and immunoassays for benzo[a]pyrene-diol-epoxide-I-modified DNA. 1988

R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
Cancer Center, School of Public Health, Columbia University, New York, NY 10032.

An interlaboratory comparison of immunoassays using antisera elicited against benzo[a]pyrene-diol-epoxide-modified DNA (BPDE-I-DNA) was carried out resulting in standardization of antisera, competitors and assay conditions. The assays used included competitive enzyme-linked immunosorbent assays (ELISA) with color and fluorescence endpoint detection and an ultrasensitive enzyme radioimmunoassay (USERIA) with a radioactive endpoint. Three different antisera were compared, two of which were obtained from different rabbits immunized with the same BPDE-I-DNA and a third from an animal immunized with another BPDE-I-DNA sample. Samples of standardized BPDE-I-DNA with high (36 pmol adduct/microgram DNA; 1.2 adducts/10(2) nucleotides) and low (4.5 fmol/microgram DNA; 1.5 adducts/10(6) nucleotides) modification levels were prepared and used in each laboratory. The antisera were all elicited against DNAs modified to a high extent, and it was therefore not surprising that they detected adducts in a slightly modified DNA sample with lower efficiency than those in highly modified DNA samples. The discrepancy of antibody recognition between the highly and slightly modified samples varied between 1.4- and 11.2-fold depending on the antiserum and assay. To ascertain the quantitative capability of the immunoassays, the modification level of DNA isolated from mouse keratinocytes treated with [3H]benzo[a]pyrene was determined by radioactivity and immunoassay. These results indicated that when a biological sample is assayed against a BPDE-I-DNA standard modified in the same range as the biological samples (4.5 fmol/microgram), quantitative recovery of adducts is achieved by immunoassay. These studies resulted in the realization that interlaboratory differences in immunoassay procedure can have significant consequences for data comparison and that where possible it is preferable for laboratories to use the same antisera and modified DNA standards.

UI MeSH Term Description Entries
D007118 Immunoassay A technique using antibodies for identifying or quantifying a substance. Usually the substance being studied serves as antigen both in antibody production and in measurement of antibody by the test substance. Immunochromatographic Assay,Assay, Immunochromatographic,Assays, Immunochromatographic,Immunoassays,Immunochromatographic Assays
D004101 Dihydroxydihydrobenzopyrenes Benzopyrenes saturated in any two adjacent positions and substituted with two hydroxyl groups in any position. The majority of these compounds have carcinogenic or mutagenic activity. Benzopyrene Dihydrodiols,Dihydrobenzopyrene Diols,Dihydrodiolbenzopyrenes,Dihydrodiols, Benzopyrene,Diols, Dihydrobenzopyrene
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D004249 DNA Damage Injuries to DNA that introduce deviations from its normal, intact structure and which may, if left unrepaired, result in a MUTATION or a block of DNA REPLICATION. These deviations may be caused by physical or chemical agents and occur by natural or unnatural, introduced circumstances. They include the introduction of illegitimate bases during replication or by deamination or other modification of bases; the loss of a base from the DNA backbone leaving an abasic site; single-strand breaks; double strand breaks; and intrastrand (PYRIMIDINE DIMERS) or interstrand crosslinking. Damage can often be repaired (DNA REPAIR). If the damage is extensive, it can induce APOPTOSIS. DNA Injury,DNA Lesion,DNA Lesions,Genotoxic Stress,Stress, Genotoxic,Injury, DNA,DNA Injuries
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000918 Antibody Specificity The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site. Antibody Specificities,Specificities, Antibody,Specificity, Antibody
D012867 Skin The outer covering of the body that protects it from the environment. It is composed of the DERMIS and the EPIDERMIS.
D015123 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide 7,8,8a,9a-Tetrahydrobenzo(10,11)chryseno (3,4-b)oxirene-7,8-diol. A benzopyrene derivative with carcinogenic and mutagenic activity. 7,8-Dihydroxy-9,10-Epoxy-7,8,9,10-Tetrahydrobenzo(a)pyrene,Benzo(a)pyrene 7,8-Dihydrodiol 9,10-Epoxide,7,8-BaP-9,10-Diol Epoxide,Anti-BaPDE,BPDE,Benzo(a)pyrene-7,8-diol 9,10-Epoxide,Anti BaPDE
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus
D018736 DNA Adducts The products of chemical reactions that result in the addition of extraneous chemical groups to DNA. DNA Adduct,Adduct, DNA,Adducts, DNA

Related Publications

R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
Monoclonal antibodies to DNA modified by a benzo[a]pyrene diol epoxide.
March 1984, Carcinogenesis,
R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
Benzo[a]pyrene diol epoxide I modification of DNA in human skin xenografts.
October 1988, The Journal of investigative dermatology,
R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
Inhibition of DNA methyltransferases in vitro by benzo[a]pyrene diol epoxide-modified substrates.
August 1984, The Journal of biological chemistry,
R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
Benzo[alpha]pyrene diol epoxide I binds to DNA at replication forks.
April 1988, Proceedings of the National Academy of Sciences of the United States of America,
R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
Benzo[a]pyrene diol-epoxide-I-DNA and oxidative DNA adducts associated with gastric adenocarcinoma.
March 1998, Cancer letters,
R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
Preferred sites of strand scission in DNA modified by andi-diol epoxide of benzo[a]pyrene.
August 1980, Science (New York, N.Y.),
R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
Detection of benzo[a]pyrene diol epoxide-DNA adducts in human placenta.
December 1988, Proceedings of the National Academy of Sciences of the United States of America,
R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
DNA adduct formation in mouse tissues in relation to serum levels of benzo(a)pyrene-diol-epoxide after injection of benzo(a)pyrene or the diol-epoxide.
February 1990, Cancer research,
R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
Identification of benzo(a)pyrene diol epoxide-binding DNA fragments using DNA immunoprecipitation technique.
April 2003, Cancer research,
R M Santella, and A Weston, and F P Perera, and G T Trivers, and C C Harris, and T L Young, and D Nguyen, and B M Lee, and M C Poirier
Topical treatment of mice with benzo[a]pyrene or parenteral administration of benzo[a]pyrene diol epoxide-DNA to rats results in faecal excretion of a putative benzo[a]pyrene diol epoxide-deoxyguanosine adduct.
September 1987, Carcinogenesis,
Copied contents to your clipboard!