NickelMaghrebV1, PascalFrancis, Checkpoint, bibRecord, 000184

Isolation and characterization of human neutralizing antibodies to rabies virus derived from a recombinant immune antibody library

Identifieur interne : 000184 ( PascalFrancis/Checkpoint ); précédent : 000183; suivant : 000185

Isolation and characterization of human neutralizing antibodies to rabies virus derived from a recombinant immune antibody library

Auteurs : Mehdi Houimel [Tunisie] ; Koussay Dellagi [Tunisie]

Source :

Journal of virological methods [ 0166-0934 ] ; 2009.

RBID : Pascal:09-0419589

Descripteurs français

Pascal (Inist)
- Homme, Virus rage, Isolement, Anticorps neutralisant, Virus recombinant, Technique phage display, Fragment peptidique Fab, Neutralisation, Microbiologie, Méthode.
Wicri :
- topic : Homme.

English descriptors

KwdEn :
- Fab-Fragment, Human, Isolation, Method, Microbiology, Neutralization, Neutralizing antibody, Phage display, Rabies virus, Recombinant virus.

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 10⁷ 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 × 10^-8 M. The Fab fragments showed dose-dependent neutralization properties for the challenge virus standard (CVS-11).

Affiliations:

Tunisie

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Le document en format XML

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<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<sup>7</sup>
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 range 7 x 10<sup>-9</sup>
 to 5 × 10<sup>-8</sup>
 M. The Fab fragments showed dose-dependent neutralization properties for the challenge virus standard (CVS-11).</div>
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Isolation and characterization of human neutralizing antibodies to rabies virus derived from a recombinant immune antibody library

Isolation and characterization of human neutralizing antibodies to rabies virus derived from a recombinant immune antibody library

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Links to Exploration step

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri