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Preparation of a monoclonal antibody specific for 5-methyl-2'-deoxycytidine and its application for the detection of DNA methylation levels in human peripheral blood cells
- Source :
- Biologicalpharmaceutical bulletin. 19(12)
- Publication Year :
- 1996
-
Abstract
- A monoclonal antibody specific for a modified nucleoside, 5-methyl-2'-deoxycytidine (m5dCyd), was prepared using 5-methylcytidine (m5Cyd)-keyhole limpet haemocyanin (KLH) conjugate, and was characterized. Termed FMC9, the antibody reacts with m5dCyd and slightly with m5Cyd and 5-methylcytosine (m5Cyt) but not with other nucleosides tested in this investigation. FMC-9 was used in an enzyme-linked immunosorbent assay (ELISA) system for the quantitation of m5dCyd levels. Sensitivity was in the picomole range. Methylation levels in peripheral blood cells of healthy donors were determined by inhibition ELISA. The percentage of m5dCyd in peripheral blood cells of 10 healthy donors was 5.08 +/- 0.50%. These results suggest that the inhibition ELISA using FMC9 is useful to monitor m5dCyd levels in the peripheral blood cells.
- Subjects :
- Adult
Male
medicine.drug_class
Ratón
Pharmaceutical Science
Enzyme-Linked Immunosorbent Assay
In Vitro Techniques
Monoclonal antibody
Deoxycytidine
chemistry.chemical_compound
Mice
Antibody Specificity
medicine
Animals
Humans
Pharmacology
Mice, Inbred BALB C
Blood Cells
Hybridomas
biology
Antibodies, Monoclonal
General Medicine
Methylation
DNA Methylation
Molecular biology
Biochemistry
chemistry
DNA methylation
Calibration
Hemocyanins
biology.protein
Female
Indicators and Reagents
Antibody
DNA
Conjugate
Subjects
Details
- ISSN :
- 09186158
- Volume :
- 19
- Issue :
- 12
- Database :
- OpenAIRE
- Journal :
- Biologicalpharmaceutical bulletin
- Accession number :
- edsair.doi.dedup.....eb5075f451326672d3335d56df5c0035