Generation of Sequence-specific, High Affinity Anti-DNA Antibodies

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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...

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Autores principales:	Cerutti, M.L., Centeno, J.M., Goldbaum, F.A., De Prat-Gay, G.
Formato:	Artículo publishedVersion
Lenguaje:	Inglés
Publicado:	2001
Materias:	Antigens Chemical bonds Complexation Dissociation DNA DNA sequences Spectroscopy Dissociation constants Antibodies DNA antibody oligonucleotide transcription factor E2 unclassified drug DNA binding protein E2 protein, Bovine papillomavirus E2 protein, Human papillomavirus type 16 monoclonal antibody oncoprotein virus protein antibody specificity antigen binding antigen recognition article binding affinity binding assay binding site carboxy terminal sequence complex formation dissociation constant DNA binding DNA sequence immunogenicity priority journal protein DNA binding protein DNA interaction spectroscopy transcription regulation amino acid sequence animal antibody combining site cattle chemistry enzyme linked immunosorbent assay human immunoglobulin variable region immunology metabolism molecular genetics mouse Papilloma virus sequence alignment sequence homology Bovinae Human papillomavirus Human papillomavirus type 16 Papillomavirus Amino Acid Sequence Animals Antibodies, Monoclonal Binding Sites Binding Sites, Antibody Cattle DNA-Binding Proteins Enzyme-Linked Immunosorbent Assay Humans Immunoglobulin Variable Region Mice Molecular Sequence Data Oncogene Proteins, Viral Papillomaviridae Sequence Alignment Sequence Homology, Amino Acid Viral Proteins
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_00219258_v276_n16_p12769_Cerutti
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	paperaa:paper_00219258_v276_n16_p12769_Cerutti2023-06-12T16:42:48Z Generation of Sequence-specific, High Affinity Anti-DNA Antibodies J. Biol. Chem. 2001;276(16):12769-12773 Cerutti, M.L. Centeno, J.M. Goldbaum, F.A. De Prat-Gay, G. Antigens Chemical bonds Complexation Dissociation DNA DNA sequences Spectroscopy Dissociation constants Antibodies DNA DNA antibody oligonucleotide transcription factor E2 unclassified drug DNA DNA binding protein E2 protein, Bovine papillomavirus E2 protein, Human papillomavirus type 16 monoclonal antibody oncoprotein virus protein antibody specificity antigen binding antigen recognition article binding affinity binding assay binding site carboxy terminal sequence complex formation dissociation constant DNA binding DNA sequence immunogenicity priority journal protein DNA binding protein DNA interaction spectroscopy transcription regulation amino acid sequence animal antibody combining site cattle chemistry enzyme linked immunosorbent assay human immunoglobulin variable region immunology metabolism molecular genetics mouse Papilloma virus sequence alignment sequence homology Bovinae Human papillomavirus Human papillomavirus type 16 Papillomavirus Amino Acid Sequence Animals Antibodies, Monoclonal Binding Sites Binding Sites, Antibody Cattle DNA DNA-Binding Proteins Enzyme-Linked Immunosorbent Assay Humans Immunoglobulin Variable Region Mice Molecular Sequence Data Oncogene Proteins, Viral Papillomaviridae Sequence Alignment Sequence Homology, Amino Acid Viral Proteins 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. 2001 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00219258_v276_n16_p12769_Cerutti
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
language	Inglés
orig_language_str_mv	eng
topic	Antigens Chemical bonds Complexation Dissociation DNA DNA sequences Spectroscopy Dissociation constants Antibodies DNA DNA antibody oligonucleotide transcription factor E2 unclassified drug DNA DNA binding protein E2 protein, Bovine papillomavirus E2 protein, Human papillomavirus type 16 monoclonal antibody oncoprotein virus protein antibody specificity antigen binding antigen recognition article binding affinity binding assay binding site carboxy terminal sequence complex formation dissociation constant DNA binding DNA sequence immunogenicity priority journal protein DNA binding protein DNA interaction spectroscopy transcription regulation amino acid sequence animal antibody combining site cattle chemistry enzyme linked immunosorbent assay human immunoglobulin variable region immunology metabolism molecular genetics mouse Papilloma virus sequence alignment sequence homology Bovinae Human papillomavirus Human papillomavirus type 16 Papillomavirus Amino Acid Sequence Animals Antibodies, Monoclonal Binding Sites Binding Sites, Antibody Cattle DNA DNA-Binding Proteins Enzyme-Linked Immunosorbent Assay Humans Immunoglobulin Variable Region Mice Molecular Sequence Data Oncogene Proteins, Viral Papillomaviridae Sequence Alignment Sequence Homology, Amino Acid Viral Proteins
spellingShingle	Antigens Chemical bonds Complexation Dissociation DNA DNA sequences Spectroscopy Dissociation constants Antibodies DNA DNA antibody oligonucleotide transcription factor E2 unclassified drug DNA DNA binding protein E2 protein, Bovine papillomavirus E2 protein, Human papillomavirus type 16 monoclonal antibody oncoprotein virus protein antibody specificity antigen binding antigen recognition article binding affinity binding assay binding site carboxy terminal sequence complex formation dissociation constant DNA binding DNA sequence immunogenicity priority journal protein DNA binding protein DNA interaction spectroscopy transcription regulation amino acid sequence animal antibody combining site cattle chemistry enzyme linked immunosorbent assay human immunoglobulin variable region immunology metabolism molecular genetics mouse Papilloma virus sequence alignment sequence homology Bovinae Human papillomavirus Human papillomavirus type 16 Papillomavirus Amino Acid Sequence Animals Antibodies, Monoclonal Binding Sites Binding Sites, Antibody Cattle DNA DNA-Binding Proteins Enzyme-Linked Immunosorbent Assay Humans Immunoglobulin Variable Region Mice Molecular Sequence Data Oncogene Proteins, Viral Papillomaviridae Sequence Alignment Sequence Homology, Amino Acid Viral Proteins Cerutti, M.L. Centeno, J.M. Goldbaum, F.A. De Prat-Gay, G. Generation of Sequence-specific, High Affinity Anti-DNA Antibodies
topic_facet	Antigens Chemical bonds Complexation Dissociation DNA DNA sequences Spectroscopy Dissociation constants Antibodies DNA DNA antibody oligonucleotide transcription factor E2 unclassified drug DNA DNA binding protein E2 protein, Bovine papillomavirus E2 protein, Human papillomavirus type 16 monoclonal antibody oncoprotein virus protein antibody specificity antigen binding antigen recognition article binding affinity binding assay binding site carboxy terminal sequence complex formation dissociation constant DNA binding DNA sequence immunogenicity priority journal protein DNA binding protein DNA interaction spectroscopy transcription regulation amino acid sequence animal antibody combining site cattle chemistry enzyme linked immunosorbent assay human immunoglobulin variable region immunology metabolism molecular genetics mouse Papilloma virus sequence alignment sequence homology Bovinae Human papillomavirus Human papillomavirus type 16 Papillomavirus Amino Acid Sequence Animals Antibodies, Monoclonal Binding Sites Binding Sites, Antibody Cattle DNA DNA-Binding Proteins Enzyme-Linked Immunosorbent Assay Humans Immunoglobulin Variable Region Mice Molecular Sequence Data Oncogene Proteins, Viral Papillomaviridae Sequence Alignment Sequence Homology, Amino Acid Viral Proteins
description	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.
format	Artículo Artículo publishedVersion
author	Cerutti, M.L. Centeno, J.M. Goldbaum, F.A. De Prat-Gay, G.
author_facet	Cerutti, M.L. Centeno, J.M. Goldbaum, F.A. De Prat-Gay, G.
author_sort	Cerutti, M.L.
title	Generation of Sequence-specific, High Affinity Anti-DNA Antibodies
title_short	Generation of Sequence-specific, High Affinity Anti-DNA Antibodies
title_full	Generation of Sequence-specific, High Affinity Anti-DNA Antibodies
title_fullStr	Generation of Sequence-specific, High Affinity Anti-DNA Antibodies
title_full_unstemmed	Generation of Sequence-specific, High Affinity Anti-DNA Antibodies
title_sort	generation of sequence-specific, high affinity anti-dna antibodies
publishDate	2001
url	http://hdl.handle.net/20.500.12110/paper_00219258_v276_n16_p12769_Cerutti
work_keys_str_mv	AT ceruttiml generationofsequencespecifichighaffinityantidnaantibodies AT centenojm generationofsequencespecifichighaffinityantidnaantibodies AT goldbaumfa generationofsequencespecifichighaffinityantidnaantibodies AT depratgayg generationofsequencespecifichighaffinityantidnaantibodies
_version_	1769810080722583552

Generation of Sequence-specific, High Affinity Anti-DNA Antibodies

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