Generation of monospecific peptide antibodies to the DNA binding domain of p53.
The DNA binding domain (DBD) is the most mutated region of p53 in tumors and has proven to be relatively resistant to the generation of specific antibodies. Template assembled synthetic peptide (TASP) synthesis of a peptide derived from the DBD creates a highly immunogenic molecule without the need for large carriers such as keyhole limpet hemocyanin (KLH). In addition, a rapid means of generating monoclonal antibodies can be achieved through immunization in conjunction with ABL/MYC retrovirus injection into recipient mice. In this paper, we demonstrate that an antibody generated by this means, KH2, reacts specifically with the DBD of p53. To date, this is the first example of a peptide immunogen used successfully in ABL/MYC monoclonal antibody production. KH2 is also the first example of a monospecific antibody that directly binds to and, by definition, assumes the conformation of the DNA binding region of p53.
Huppi
K
K
Laboratory of Genetics, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. huppi@helix.nih.gov
Henderson
D
D
Siwarski
D
D
Hochman
J
J
Bergel
M
M
Tuchscherer
G
G
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D000918:Antibody Specificity / Q000276:immunology*; D001665:Binding Sites; D015153:Blotting, Western; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain / Q000235:genetics; D016313:Genes, abl; D016259:Genes, myc; D006801:Humans; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D010954:Plasmacytoma / Q000235:genetics / Q000276:immunology; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D016159:Tumor Suppressor Protein p53