Isolation and characterization of human neutralizing antibodies to rabies virus derived from a recombinant immune antibody library.
Identifieur interne : 000083 ( Main/Exploration ); précédent : 000082; suivant : 000084Isolation and characterization of human neutralizing antibodies to rabies virus derived from a recombinant immune antibody library.
Auteurs : Mehdi Houimel [Tunisie] ; Koussay DellagiSource :
- Journal of virological methods [ 1879-0984 ] ; 2009.
Descripteurs français
- KwdFr :
- ARN (génétique), ARN (isolement et purification), Anticorps antiviraux (génétique), Anticorps antiviraux (immunologie), Anticorps antiviraux (isolement et purification), Banque de peptides (MeSH), Cartographie épitopique (MeSH), Femelle (MeSH), Fragments Fab d'immunoglobuline (génétique), Fragments Fab d'immunoglobuline (immunologie), Fragments Fab d'immunoglobuline (isolement et purification), Glycoprotéines (immunologie), Humains (MeSH), Lymphocytes (immunologie), Mâle (MeSH), Protéines virales (immunologie), Rage (maladie) (génétique), Rage (maladie) (immunologie), Région variable d'immunoglobuline (génétique), Région variable d'immunoglobuline (immunologie), Région variable d'immunoglobuline (isolement et purification), Tests de neutralisation (MeSH), Vaccins antirabiques (immunologie), Virus de la rage (immunologie).
- MESH :
- génétique : ARN, Anticorps antiviraux, Fragments Fab d'immunoglobuline, Rage (maladie), Région variable d'immunoglobuline.
- immunologie : Anticorps antiviraux, Fragments Fab d'immunoglobuline, Glycoprotéines, Lymphocytes, Protéines virales, Rage (maladie), Région variable d'immunoglobuline, Vaccins antirabiques, Virus de la rage.
- isolement et purification : ARN, Anticorps antiviraux, Fragments Fab d'immunoglobuline, Région variable d'immunoglobuline.
- Banque de peptides, Cartographie épitopique, Femelle, Humains, Mâle, Tests de neutralisation.
English descriptors
- KwdEn :
- Antibodies, Viral (genetics), Antibodies, Viral (immunology), Antibodies, Viral (isolation & purification), Epitope Mapping (MeSH), Female (MeSH), Glycoproteins (immunology), Humans (MeSH), Immunoglobulin Fab Fragments (genetics), Immunoglobulin Fab Fragments (immunology), Immunoglobulin Fab Fragments (isolation & purification), Immunoglobulin Variable Region (genetics), Immunoglobulin Variable Region (immunology), Immunoglobulin Variable Region (isolation & purification), Lymphocytes (immunology), Male (MeSH), Neutralization Tests (MeSH), Peptide Library (MeSH), RNA (genetics), RNA (isolation & purification), Rabies (genetics), Rabies (immunology), Rabies Vaccines (immunology), Rabies virus (immunology), Viral Proteins (immunology).
- MESH :
- chemical , genetics : Antibodies, Viral, Immunoglobulin Fab Fragments, Immunoglobulin Variable Region, RNA.
- chemical , immunology : Antibodies, Viral, Glycoproteins, Immunoglobulin Fab Fragments, Immunoglobulin Variable Region, Rabies Vaccines, Viral Proteins.
- chemical , isolation & purification : Antibodies, Viral, Immunoglobulin Fab Fragments, Immunoglobulin Variable Region, RNA.
- genetics : Rabies.
- immunology : Lymphocytes, Rabies, Rabies virus.
- Epitope Mapping, Female, Humans, Male, Neutralization Tests, Peptide Library.
Abstract
A human immune Fab library was constructed using RNAs from peripheral blood lymphocytes obtained from rabies virus hyperimmune volunteers on phagemid vector. The size of the constructed Fab library was 2 x 10(7) Escherichia coli transformants. After four rounds of panning on whole inactivated rabies virus (PV-11), phage clones displaying rabies virus-specific human Fab were selected. The specificity of soluble Fab antibody fragments, derived from positive phage clones was verified by ELISA. Among 20 specific Fab clones, the genetic sequence of 6 of them (FabRV01, FabRV02, FabRV03, FabRV04, FabRV05, and FabRV06) was analyzed. The variable heavy (VH) and variable light (VL) domains were found to share 90% and 93% homology with sequences encoded by the corresponding human germline genes, respectively. The soluble Fab fragments, expressed in Escherichia coli were purified by a single step Nickel-NTA affinity chromatography via a hexa-histidine tag and their binding specificities to rabies virus were confirmed. Three of the Fab antibodies, FabRV01, FabRV02 and FabRV03, showed binding characteristics to rabies virus glycoprotein antigenic site III with affinities in the K(D) range 7 x 10(-9) to 5 x 10(-8)M. The Fab fragments showed dose-dependent neutralization properties for the challenge virus standard (CVS-11).
DOI: 10.1016/j.jviromet.2009.06.018
PubMed: 19559727
Affiliations:
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Le document en format XML
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(immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN (génétique)</term><term>ARN (isolement et purification)</term><term>Anticorps antiviraux (génétique)</term><term>Anticorps antiviraux (immunologie)</term><term>Anticorps antiviraux (isolement et purification)</term><term>Banque de peptides (MeSH)</term><term>Cartographie épitopique (MeSH)</term><term>Femelle (MeSH)</term><term>Fragments Fab d'immunoglobuline (génétique)</term><term>Fragments Fab d'immunoglobuline (immunologie)</term><term>Fragments Fab d'immunoglobuline (isolement et purification)</term><term>Glycoprotéines (immunologie)</term><term>Humains (MeSH)</term><term>Lymphocytes (immunologie)</term><term>Mâle (MeSH)</term><term>Protéines virales (immunologie)</term><term>Rage (maladie) (génétique)</term><term>Rage (maladie) (immunologie)</term><term>Région variable d'immunoglobuline (génétique)</term><term>Région variable d'immunoglobuline (immunologie)</term><term>Région variable d'immunoglobuline (isolement et purification)</term><term>Tests de neutralisation (MeSH)</term><term>Vaccins antirabiques (immunologie)</term><term>Virus de la rage (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antibodies, Viral</term><term>Immunoglobulin Fab Fragments</term><term>Immunoglobulin Variable Region</term><term>RNA</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Viral</term><term>Glycoproteins</term><term>Immunoglobulin Fab Fragments</term><term>Immunoglobulin Variable Region</term><term>Rabies Vaccines</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Antibodies, Viral</term><term>Immunoglobulin Fab Fragments</term><term>Immunoglobulin Variable Region</term><term>RNA</term></keywords><keywords scheme="MESH" qualifier="genetics" 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d'immunoglobuline</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Epitope Mapping</term><term>Female</term><term>Humans</term><term>Male</term><term>Neutralization Tests</term><term>Peptide Library</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Banque de peptides</term><term>Cartographie épitopique</term><term>Femelle</term><term>Humains</term><term>Mâle</term><term>Tests de neutralisation</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A human immune Fab library was constructed using RNAs from peripheral blood lymphocytes obtained from rabies virus hyperimmune volunteers on phagemid vector. The size of the constructed Fab library was 2 x 10(7) Escherichia coli transformants. After four rounds of panning on whole inactivated rabies virus (PV-11), phage clones displaying rabies virus-specific human Fab were selected. The specificity of soluble Fab antibody fragments, derived from positive phage clones was verified by ELISA. Among 20 specific Fab clones, the genetic sequence of 6 of them (FabRV01, FabRV02, FabRV03, FabRV04, FabRV05, and FabRV06) was analyzed. The variable heavy (VH) and variable light (VL) domains were found to share 90% and 93% homology with sequences encoded by the corresponding human germline genes, respectively. The soluble Fab fragments, expressed in Escherichia coli were purified by a single step Nickel-NTA affinity chromatography via a hexa-histidine tag and their binding specificities to rabies virus were confirmed. Three of the Fab antibodies, FabRV01, FabRV02 and FabRV03, showed binding characteristics to rabies virus glycoprotein antigenic site III with affinities in the K(D) range 7 x 10(-9) to 5 x 10(-8)M. The Fab fragments showed dose-dependent neutralization properties for the challenge virus standard (CVS-11).</div></front></TEI><affiliations><list><country><li>Tunisie</li></country></list><tree><noCountry><name sortKey="Dellagi, Koussay" sort="Dellagi, Koussay" uniqKey="Dellagi K" first="Koussay" last="Dellagi">Koussay Dellagi</name></noCountry><country name="Tunisie"><noRegion><name sortKey="Houimel, Mehdi" sort="Houimel, Mehdi" uniqKey="Houimel M" first="Mehdi" last="Houimel">Mehdi Houimel</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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