Quantitation and visualization of alkyl deoxynucleosides in the DNA of mammalian cells by monoclonal antibodies
Monoclonal antibodies (Mab) characterized by a high substrate specificity and affinity, in combination with radio- and enzyme-immunoassays, or with immuno-slot-blot techniques, now permit the detection of femtomole to subfemtomole amounts of, e.g., alkyldeoxynucleosides in small samples of DNA isolated from tissues or cultured cells previously exposed to nonradioactive N-nitroso compounds. Furthermore, selected Mab can be used to quantitate by direct immunofluorescence specific alkyldeoxynucleosides in the nuclear DNA of single cells. With this method, the detection limit for the alkylation product O/sup 6/-ethyldeoxyguanosine (O/sup 6/-EtdGuo) is presently of the order of 10/sup 2/-10/sup 3/ O/sup 6/-EtdGuo residues per diploid mammalian genome. Individual cells can thus be monitored for the presence of specific carcinogen-DNA adducts, and with respect to their capacity for enzymatic removal of such modified structures from DNA (as exemplified here by the kinetics of the enzymatic elimination of O/sup 6/-EtdGuo from the DNa of malignant neurogenic rat cell lines). In combination with transmission electron microscopy, Mab also permit direct visualization of specific carcinogen-modified structures in individual DNA molecules. DNA strands of defined nucleotide sequence can thus be analyzed by immuno-electron microscopy for the presence of hot spots of specific structural alterations caused by defined carcinogens/mutagens.
- Research Organization:
- Universitaet Essen (GH), West Germany
- OSTI ID:
- 5460593
- Journal Information:
- Environ. Health Perspect.; (United States), Vol. 62
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
DNA ADDUCTS
ENZYME IMMUNOASSAY
RADIOIMMUNOASSAY
NUCLEOSIDES
ALKYLATION
FLUORESCENCE
MONOCLONAL ANTIBODIES
TRANSMISSION ELECTRON MICROSCOPY
ADDUCTS
ANTIBODIES
CHEMICAL REACTIONS
ELECTRON MICROSCOPY
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
LUMINESCENCE
MICROSCOPY
NUCLEOTIDES
ORGANIC COMPOUNDS
RADIOASSAY
RADIOIMMUNOLOGY
RIBOSIDES
TRACER TECHNIQUES
550601* - Medicine- Unsealed Radionuclides in Diagnostics