CovidV1, Main, Exploration, bibRecord, 000C51

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

Identifieur interne : 000C51 ( Main/Exploration ); précédent : 000C50; suivant : 000C52

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

Auteurs : Chen Li [République populaire de Chine] ; Feng Zhang [République populaire de Chine] ; Hong Lin [République populaire de Chine] ; Zhong-Can Wang [République populaire de Chine] ; Xin-Jian Liu [République populaire de Chine] ; Zhen-Qing Feng [République populaire de Chine] ; Jin Zhu [République populaire de Chine] ; Xiao-Hong Guan [République populaire de Chine]

Source :

Acta Pharmacologica Sinica [ 1671-4083 ] ; 2011.

RBID : PMC:4002766

Abstract

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.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4002766

DOI: 10.1038/aps.2010.209
PubMed: 21278782
PubMed Central: 4002766

Affiliations:

République populaire de Chine

Le document en format XML

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<front><div type="abstract" xml:lang="en"><sec><title>Aim:</title>
<p>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.</p>
</sec>
<sec><title>Methods:</title>
<p>The Fab gene was amplified using overlap PCR and inserted into the vector pComb3XSS. The recombinant vector was then transformed into <italic>E coli</italic>
 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 <italic>in vivo</italic>
.</p>
</sec>
<sec><title>Results:</title>
<p>A recombinant vector was constructed. The Fab was expressed in soluble form in <italic>E coli</italic>
 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 (<italic>P</italic>
<0.05, Logrank test).</p>
</sec>
<sec><title>Conclusion:</title>
<p>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.</p>
</sec>
</div>
</front>
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Generation and characterization of the human neutralizing antibody fragment Fab091 against rabies virus

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

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