Generation of Sequence-specific, High Affinity Anti-DNA Antibodies
| Autor: | Cerutti, M.L., Centeno, J.M., Goldbaum, F.A., De Prat-Gay, G. |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2001 |
| Předmět: |
Immunoglobulin Variable Region
E2 protein Bovine papillomavirus DNA sequences immunogenicity dissociation constant immunology Mice binding affinity Human papillomavirus type 16 virus protein antigen binding animal Papillomaviridae Spectroscopy binding assay E2 protein Human papillomavirus type 16 Chemical bonds article Antibodies Monoclonal Papillomavirus unclassified drug antigen recognition DNA-Binding Proteins oncoprotein priority journal sequence alignment protein DNA interaction Complexation transcription regulation Dissociation oligonucleotide Human papillomavirus antibody combining site Molecular Sequence Data DNA sequence Enzyme-Linked Immunosorbent Assay protein DNA binding chemistry Antibodies Bovinae Viral Proteins complex formation Animals Humans DNA antibody human Amino Acid Sequence Antigens DNA binding mouse carboxy terminal sequence Binding Sites Sequence Homology Amino Acid binding site antibody specificity transcription factor E2 Papilloma virus DNA sequence homology Oncogene Proteins Viral DNA binding protein enzyme linked immunosorbent assay Dissociation constants monoclonal antibody cattle molecular genetics Binding Sites Antibody metabolism |
| Zdroj: | J. Biol. Chem. 2001;276(16):12769-12773 Biblioteca Digital (UBA-FCEN) Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales instacron:UBA-FCEN |
| Popis: | 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. Fil:Cerutti, M.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:De Prat-Gay, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. |
| Databáze: | OpenAIRE |
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