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. 1986 May 1;89(1):123-30.
doi: 10.1016/0022-1759(86)90040-2.

Characterization of monoclonal antibody to DNA.RNA and its application to immunodetection of hybrids

S J Boguslawski, D E Smith, M A Michalak, K E Mickelson, C O Yehle, W L Patterson, R J Carrico

Characterization of monoclonal antibody to DNA.RNA and its application to immunodetection of hybrids

S J Boguslawski et al. J Immunol Methods. .

Abstract

Monoclonal antibodies were raised to a DNA.RNA heteropolymer duplex prepared by transcription of phi X174 single-stranded DNA with DNA-dependent RNA polymerase. A monoclonal antibody with the highest affinity and specificity was selected. This antibody bound the DNA.RNA heteropolymer and poly(I).poly(dC) equally but 100-fold higher levels of poly(A).poly(dT) were required to achieve a similar degree of binding in competitive binding assays using DNA.[3H]RNA. Single-stranded DNA, double-stranded DNA and RNA, and ribosomal RNA were not bound by the antibody. The observed association constant for the antibody and DNA.[3H]RNA, determined by Scatchard analysis, was 8.5 X 10(10) l/mol assuming independent antibody binding sites. The antibody and an alkaline phosphatase-labeled second antibody were used in an immunodetection method for measurement of hybrids formed between immobilized DNA probes of various lengths and 23 S ribosomal RNA. The colorimetric response of this assay increased linearly with the amount of hybrid formed.

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