Natl Res Council Canada, Inst Plant Biotechnol, Saskatoon, SK S7N 0W9, Canada; Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Peoples R China; Univ Saskatchewan, Dept Microbiol & Immunol, Saskatoon, SK S7N 5E5, Canada
New approaches of making single chain Fv antibodies against O-6-methyl-2'-deoxyguanosine (O(6)MdG) have been demonstrated by using the phage antibody display system. Using O(6)MdG as an antigen, 21 positive clones were identified by ELISA from this library, one of which, designated H3, specifically binds to O(6)MdG with high affinity. The H3 scFv antibody has an affinity constant (K-aff) of 5.94 x 10(11)(mol/L)(-1). H3 scFv has been successfully used to detect O-6 MdG in DNA hydrolyses from yeast or E. coli cells treated with a DNA methylating agent. To our knowledge, this is the first report of the selection of a specific scFv against DNA adducts. The results demonstrate the potential applications of the phage display technology for the detection of DNA lesions caused by mutagens and carcinogens.
DAI Heping; ZHAO Rouhong; Haug Brenda; Sean M. Hemmingsen; XIAO Wei.Isolation by phage display and characterization of a single-chain antibody specific for O-6-methyldeoxyguanosine,CHINESE SCIENCE BULLETIN,2001,46(12):1024-1029