NUCLEIC ACID-REACTIVE ANTIBODIES SPECIFIC FOR NUCLEOSIDES AND NUCLEOTIDES

Abstract

ABSTRACT A method is described for making anti-purine and anti-pyrimidine anti-bodies by immunization with conjugates of naturally-occurring nucleosides or nucleotides with carrier proteins. The specificities of the antisera are presented and shown to be predominantly, if not solely, for the determinant purine or pyrimidine group presented in the antigen. The antisera react with single-stranded or denatured DNA and, in the case of anti-adenosine, with RNA preparations. The antisera have been utilized in biochemical systems to inhibit the priming ability of DNA in a DNA-dependent DNA polymerase system, to detect minor bases such as 6-methyladenosine in DNA, and to detect small areas of single-strandedness in otherwise native DNA. The sera have also been shown to react with living cells, inhibiting the development of fertilized sea urchin eggs and entering and inhibiting transformed Chinese hamster lung cells without affecting normal ones. They have been used in immunofluorescence experiments in which they have been shown to react with nuclear DNA of fixed mouse L-cells harvested during the S phase. Moreover, they have found application in the characterization and identification of human and mouse chromosomes. Finally, initial studies have shown that they can be used in highly specific radioimmunoassays for purine and pyrimidine nucleosides.