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Artículo

Generation of sequence-specific, high affinity anti-DNA antibodies.

Cerutti, Maria LauraIcon ; Centeno, Juan M.; Goldbaum, Fernando AlbertoIcon ; de Prat Gay, GonzaloIcon
Fecha de publicación: 12/2001
Editorial: American Society for Biochemistry and Molecular Biology
Revista: Journal of Biological Chemistry (online)
ISSN: 0021-9258
e-ISSN: 1083-351X
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Biofísica; Bioquímica y Biología Molecular

Resumen

By taking advantage of the extreme stability of a protein-DNA complex, we have obtained two highly specific monoclonal antibodies against a predetermined palindromic DNA sequence corresponding to the binding site of the E2 transcriptional regulator of the human papillomavirus (HPV-16). The purified univalent antibody fragments bind to a double-stranded DNA oligonucleotide corresponding to the E2 binding site in solution with dissociation constants in the low and subnanomolar range. This affinity matches that of the natural DNA binding domain and is severalfold higher than the affinity of a homologous bovine E2 C-terminal domain (BPV-1) for the same DNA. These antibodies discriminate effectively among a number of double- and single-stranded synthetic DNAs with factors ranging from 125- to 20,000-fold the dissociation constant of the specific DNA sequence used in the immunogenic protein-DNA complex. Moreover, they are capable of fine specificity tuning, since they both bind less tightly to another HPV-16 E2 binding site, differing in only 1 base pair in a noncontact flexible region. Beyond the relevance of obtaining a specific anti-DNA response, these results provide a first glance at how DNA as an antigen is recognized specifically by an antibody. The accuracy of the spectroscopic method used for the binding analysis suggests that a detailed mechanistic analysis is attainable.
Palabras clave: Anti-Dna , Affinity , Binding , Monoclonal Antibody
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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)
Identificadores
URI: http://hdl.handle.net/11336/48130
URL: http://www.jbc.org/content/276/16/12769.long
DOI: http://dx.doi.org/10.1074/jbc.M100260200
Colecciones
Articulos(IIBBA)
Articulos de INST.DE INVEST.BIOQUIMICAS DE BS.AS(I)
Citación
Cerutti, Maria Laura; Centeno, Juan M.; Goldbaum, Fernando Alberto; de Prat Gay, Gonzalo; Generation of sequence-specific, high affinity anti-DNA antibodies.; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 276; 16; 12-2001; 12769-12773
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