Isolation by phage display and characterization of a single-chain antibody specific for O^6-methyldeoxyguanosine  

作　　者：DAI Heping ZHAO Rouhong Haug Brenda Sean M.Hemmingsen XIAO Wei 

机构地区：[1]Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan 430072,China [2]Plant Biotechnology Institute,National Research Council of Canada,Saskatoon,SK S7N OW9,Canada [3]Department of Microbiology and Immunology,University of Saskatchewan,Saskatoon,SK S7N 5E5,Canada

出　　处：《Chinese Science Bulletin》2001年第12期1024-1029,共6页

基　　金：This work was supported by the President Grant of the Chinese Academy of Sciences and K.C. Wong Education Founda-tion (Hong Kong) to HD., and also by the Natural Sciences and Engineering Research Council of Canada (Grant No. OGPO 138338 to WX).

摘　　要：New approaches of making single chain Fv antibodies against O6-methyl-2’ -deoxyguanosine (O6MdG) have been demonstrated by using the phage antibody display system. Using O6MdG as an antigen, 21 positive clones were identified by ELISA from this library, one of which, designated H3, specifically binds to O6MdG with high affinity. The H3 scFv antibody has an affinity constant (Kaff) of 5.94×1011(mol/L)-1. H3 scFv has been successfully used to detect O6MdG 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.New approaches of making single chain Fv antibodies against O6-methyl-2 ′-deoxyguanosine (O6MdG) have been demonstrated by using the phage antibody display system. Using O6MdG as an antigen, 21 positive clones were identified by ELISA from this library, one of which, designated H3, specifically binds to O6MdG with high affinity. The H3 scFv antibody has an affinity constant (Kaff) of 5.94x1011(mol/L)?1. H3 scFv has been successfully used to detect O6MdG 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.

关 键 词：O6-methyl-2’ -deoxyguanosine  PHAGE display antibody  single chain FV DNA adducts. 

分 类 号：R392[医药卫生—免疫学]