Measurement of ultraviolet light-induced photolesions in mammalian DNA by microELISA

A sensitive microELISA was developed for titration of anti-UV-DNA antisera and measurement of photolesions in DNA using UV-irradiated DNA as antigen coupled to the wells of microtitre plates. Efficient and reproducible immobilization of the antigen required precoating of the plates with poly(L-lysine). Antisera from 2 rabbits immunized with UVdsDNA recognized specifically thymine dimer associated lesions. The antiserum from one animal was used to probe photodamage in DNA. Lesions induced by an irradiation dose as small as 2.5 J/m2 could be detected.