Anti-8-ore-2 '-deoxyguanosine phage antibodies: Isolation, characterization, and relationship to disease states

We have used human single chain Fv (scFv) phage display antibody libraries to isolate recombinant antibodies against the DNA adduct 8-oxi-2'-deoxyguan osine (8-oxodG). One of these scFvs (175G) bound to several 8-oxodG-contain ing oligonucleotides whilst demonstrating no cross-reactivity with G-contai ning control oligonucleotides, and bound to 8-oxodG; lesions introduced int o DNA by treatment with methylene blue and white Light. In addition, 175G i nhibited the cleavage of an 8-oxodG-containing oligonucleotide by the Esche richia coli enzyme formamidopyrimidine-DNA glycosylase (Fpg). The nucleotid e sequence of the 1756 V-H gene segment was 98% homologous to the published V-H sequence of a human hybridoma derived from a patient with systemic lup us erythematosus (SLE). Sera from two SLE patients bound to damaged DNA, an d this binding could be inhibited by 175G. The use of human scFv phage disp lay libraries has thus produced a unique reagent with specificity for 8-oxo dG which may have a role in damage detection and quantitation and in modify ing DNA repair activity. 175G also offers support to the hypothesis that SL E might be associated with oxidative damage to DNA (C) 2001 Academic Press.