Preparation and characterization of an anti-DNA monoclonal antibody showing size selectivity toward DNA fragments.
Anti-DNA monoclonal antibodies were prepared using an in vitro immunization method. Balb/c mouse splenocytes were immunized with HeLa cell nuclear extract in the presence of N-acetylmuramyl-L-alanyl-D-isoglutamine and fused with P3U1 myeloma cells using PEG 4000. After HAT selection and ELISA using fragmented HeLa genomic DNA, an anti-DNA monoclonal antibody was obtained. The monoclonal antibody D-1-1, whose isotype was IgM, interacted with a variety of double-stranded DNA. The antibody reacted only with DNA fragments longer than 0.8 kbp, and its apparent dissociation constant for a 1.0-kbp DNA fragment was 34 nM. This antibody will be a helpful tool for the detection of DNA structures.
Onishi
Yoshiaki
Y
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Ibaraki, Japan. y-onishi@aist.go.jp
Kato
Megumi
M
Hanyu
Yoshiro
Y
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D015394:Molecular Structure; D011485:Protein Binding / Q000276:immunology
D000911:Antibodies, Monoclonal; D004247:DNA