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Monoclonal antipeptide antibodies: Affinity and kinetic rate constants measured for the peptide and the cognate protein using a biosensor technology

Identifieur interne : 00DA82 ( Main/Exploration ); précédent : 00DA81; suivant : 00DA83

Monoclonal antipeptide antibodies: Affinity and kinetic rate constants measured for the peptide and the cognate protein using a biosensor technology

Auteurs : Gabrielle Zeder-Lutz [France] ; Daniéle Altschuh [France] ; H. Mario Geysen [Australie] ; Elisabeth Trifilieff [France] ; Ghislaine Sommermeyer [France] ; Marc H. V. Van Regenmortel [France]

Source :

RBID : ISTEX:DC1B06FEF1D4ED26B8553BC5DCF6A953D2816C77

English descriptors

Abstract

Abstract: The interaction of antipeptide antibodies with the corresponding peptide and the cognate protein has been compared using a novel biosensor technology (BIAcoreTM Pharmacia). The peptide corresponds to residues 110–135 of the coat protein of tobacco mosaic virus, known to encompass an α-helical region reactive with antiprotein antibodies. A panel of 33 monoclonal antibodies raised against the peptide was studied and the epitope recognized by these antibodies was determined by the pepscan method. Further discrimination between the antibodies was performed by measurements of association and dissociation kinetic constants. Several antibodies showed an heterogeneous binding profile when reacting with the 25 residue long peptide but not with a shorter 10 residue peptide suggesting that they recognized different conformational states in the longer peptide. Equilibrium affinity constants were calculated for five of the antibodies and were found to be 10–50 times higher for the peptide than for the protein, the difference being caused mainly by a lower association rate constant.

Url:
DOI: 10.1016/0161-5890(93)90086-Q


Affiliations:


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Le document en format XML

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<term>Antibody</term>
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<term>Antipeptide antibodies</term>
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<term>Antiprotein antibodies</term>
<term>Ascites fluid</term>
<term>Background binding</term>
<term>Biacore</term>
<term>Biacore experiments</term>
<term>Biacore system</term>
<term>Biacoretm system</term>
<term>Biosensor</term>
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<term>Dissociation rate constants</term>
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<term>Equilibrium affinity constants</term>
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<term>Flexible keys</term>
<term>Free peptide</term>
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<term>High frequency</term>
<term>Homogeneous binding</term>
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<term>Immunization</term>
<term>Immunizing peptide</term>
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<term>Kinetic rate constants</term>
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<term>Matrix</term>
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<term>Monoclonal antibodies</term>
<term>Monoclonal antibody</term>
<term>Monoclonal antipeptide antibodies</term>
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<term>Negative control</term>
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<term>Pepscan</term>
<term>Pepscan method</term>
<term>Peptide</term>
<term>Peptide synthesis</term>
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<term>Rate constants</term>
<term>Reaction rate</term>
<term>Regenmortel</term>
<term>Same concn</term>
<term>Second half</term>
<term>Sensor chip</term>
<term>Short peptides</term>
<term>Shorter peptide</term>
<term>Solid phase</term>
<term>Specific antibodies</term>
<term>Spleen cells</term>
<term>Standard deviation</term>
<term>Standard deviations</term>
<term>Straight line</term>
<term>Surface plasmon resonance</term>
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<term>Synthetic peptides</term>
<term>Titertek multiscan elisa reader</term>
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<term>Various peptides</term>
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<term>Adjustable locks</term>
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<term>Conformation</term>
<term>Conformational</term>
<term>Conformational adaptation</term>
<term>Conformational states</term>
<term>Data points</term>
<term>Dextran</term>
<term>Dextran matrix</term>
<term>Dissociation rate</term>
<term>Dissociation rate constants</term>
<term>Elisa</term>
<term>Epitope</term>
<term>Epitope mapping</term>
<term>Equilibrium affinity</term>
<term>Equilibrium affinity constants</term>
<term>Equivalent reactivity</term>
<term>Fine specificity</term>
<term>First half</term>
<term>Flexible keys</term>
<term>Free peptide</term>
<term>Helical conformation</term>
<term>Heterogeneous binding</term>
<term>High frequency</term>
<term>Homogeneous binding</term>
<term>Hybridoma culture supernatants</term>
<term>Immun</term>
<term>Immunization</term>
<term>Immunizing peptide</term>
<term>Independent experiments</term>
<term>Kinetic analysis</term>
<term>Kinetic constants</term>
<term>Kinetic measurements</term>
<term>Kinetic rate constants</term>
<term>Mabs</term>
<term>Matrix</term>
<term>Molec</term>
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<term>Sensor chip</term>
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<term>Shorter peptide</term>
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<term>Spleen cells</term>
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<term>Synthetic peptides</term>
<term>Titertek multiscan elisa reader</term>
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<div type="abstract" xml:lang="en">Abstract: The interaction of antipeptide antibodies with the corresponding peptide and the cognate protein has been compared using a novel biosensor technology (BIAcoreTM Pharmacia). The peptide corresponds to residues 110–135 of the coat protein of tobacco mosaic virus, known to encompass an α-helical region reactive with antiprotein antibodies. A panel of 33 monoclonal antibodies raised against the peptide was studied and the epitope recognized by these antibodies was determined by the pepscan method. Further discrimination between the antibodies was performed by measurements of association and dissociation kinetic constants. Several antibodies showed an heterogeneous binding profile when reacting with the 25 residue long peptide but not with a shorter 10 residue peptide suggesting that they recognized different conformational states in the longer peptide. Equilibrium affinity constants were calculated for five of the antibodies and were found to be 10–50 times higher for the peptide than for the protein, the difference being caused mainly by a lower association rate constant.</div>
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