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

Houimel, Mehdi; Dellagi, Koussay

doi:10.1016/j.jviromet.2009.06.018

Cited by 13 publications

(5 citation statements)

References 61 publications

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“…The antibody variable domains can be engineered into small fragments ( Table 2), including scFvs and F[ab] molecules [76][77][78] which do not require production in costly eukaryotic expression systems. Several of these small antibody fragments have been investigated regarding their antiviral activities [79,80], including camelid VHH domains. The serum of camels, dromedaries and llamas contains a unique type of antibodies devoid of antibody light chains [21].…”

Section: Antibody Engineeringmentioning

confidence: 99%

Monoclonal antibodies for prophylactic and therapeutic use against viral infections

et al. 2013

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Section: Antibody Engineeringmentioning

confidence: 99%

Monoclonal antibodies for prophylactic and therapeutic use against viral infections

et al. 2013

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“…However, a reasonable immunized library typically contains 10 7 to 10 8 independent transformants [30] and Fab-phage have been successfully isolated from immune libraries with sizes of ~10 7 (see for example refs. [31, 32]). Thus our library was deemed large enough to proceed.…”

Section: Resultsmentioning

confidence: 99%

A parallel panning scheme used for selection of a GluA4-specific Fab targeting the ligand-binding domain

Clausen

Mohr

Riise

et al. 2016

International Journal of Biological Macromolecules

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A method for development of murine Fab fragments towards extracellular domains of a surface receptor is presented. The GluA4 ionotropic glutamate receptor is used as a model system. Recombinant GluA4 ectodomain comprising both the N-terminal domain (NTD) and the ligand-binding domain (LBD) in one molecule was used for immunization. A Fab-phage library was constructed and a parallel panning approach enabled selection of murine Fab fragments towards either intact ectodomain or the isolated LBD of the GluA4 receptor. One LBD-Fab (FabL9) showed exclusive selectivity for the GluA4 LBD, over a panel of LBDs from GluA2, GluK1, GluK2 and GluD2. Soluble FabL9 was produced in amounts suitable for characterization. Competitive ELISA and rat-brain immunoprecipitation experiments confirmed that the FabL9 epitope is conserved in the LBD and in the intact native receptor. By an alignment of GluA2 and GluA4, the likely binding epitope for FabL9 was predicted. This study demonstrates a simple approach for development of antibody fragments towards specific sub-domains of a large ligand-gated ion channel, and this method could be utilized for all multi-domain surface receptors where antibody domain-selectivity may be desirable. Furthermore, we present for the first time a GluA4 subtype-specific murine Fab fragment targeting the LBD of the receptor.

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“…First, the in vivo clearance of RABV is a complicated process. Compared with small-molecular-weight antibodies, the IgG antibody (HRIG), with antibody-dependent cellular cytotoxicity and complement-mediated cytotoxicity, has a greater scavenging capacity in vivo [33,34] . The neutralizing activity of Fab is usually less than that of full-length IgG because of the monovalence of the Fab fragments and the lack of Fc fragment on an IgG molecule.…”

Section: Vaccinementioning

confidence: 99%

Generation and characterization of the human neutralizing antibody fragment Fab091 against rabies virus

Zhang

Lin

et al. 2011

Acta Pharmacol Sin

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Aim: To transform the human anti-rabies virus glycoprotein (anti-RABVG) single-chain variable fragment (scFv) into a Fab fragment and to analyze its immunological activity. Methods: The Fab gene was amplified using overlap PCR and inserted into the vector pComb3XSS. The recombinant vector was then transformed into E coli Top10F′ for expression and purification. The purified Fab was characterized using SDS-PAGE, Western blotting, indirect ELISA, competitive ELISA, and the fluorescent antibody virus neutralization test (FAVN), respectively, and examined in a Kunming mouse challenge model in vivo. Results: A recombinant vector was constructed. The Fab was expressed in soluble form in E coli Top10F′. Specific binding of the Fab to rabies virus was confirmed by indirect ELISA and immunoprecipitation (IP). The neutralizing antibody titer of Fab was 10.26 IU/mL. The mouse group treated with both vaccine and human rabies immunoglobulin (HRIG)/Fab091 (32 IU/kg) showed protection against rabies, compared with the control group (P<0.05, Logrank test). Conclusion: The antibody fragment Fab was shown to be a neutralizing antibody against RABVG. It can be used together with other monoclonal antibodies for post-exposure prophylaxis of rabies virus in future studies.

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

Cited by 13 publications

References 61 publications

Monoclonal antibodies for prophylactic and therapeutic use against viral infections

Monoclonal antibodies for prophylactic and therapeutic use against viral infections

A parallel panning scheme used for selection of a GluA4-specific Fab targeting the ligand-binding domain

Generation and characterization of the human neutralizing antibody fragment Fab091 against rabies virus

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