Detection of DNA Adducts in Human Breast Tissues
Abstract
The purpose of this project is to optimize and apply new methodology for the detection of unknown DNA adducts in breast and, for comparison, other human tissues. Current methodology for this purpose has shortcomings, and breast cancer may be initiated by unknown DNA adducts. Initially the methodology will be applied to nonpolar adducts, and later it can be expanded. The concept for the new methodology is the same as 32P-postlabeling, but chemical labeling is done with a fluorescent tag to overcome variation in labeling yields, increase resolution, and provide compatibility with mass spectrometry. As part of optimization, a prototype fluorescent tag has been characterized in terms of its structural and reactivity properties. This has led to the detection of 60 fmol of dAMP by the method. Also, to facilitate purification of the tag, and improve removal of excess tag after the labeling reaction, its separation has been studied by immobilized metal affinity chromatography. This is intended to bring the sensitivity of the method closer to the low amol level that we have reached for standards.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 1997
- Accession Number
- ADB230256
Entities
People
- Roger Giese
Organizations
- Northeastern University