Generation and characterization of the human neutralizing antibody fragment Fab091 against rabies virus
Identifieur interne : 000206 ( Ncbi/Merge ); précédent : 000205; suivant : 000207Generation 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.
Abstract
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.
The Fab gene was amplified using overlap PCR and inserted into the vector pComb3XSS. The recombinant vector was then transformed into
A recombinant vector was constructed. The Fab was expressed in soluble form in
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:
DOI: 10.1038/aps.2010.209
PubMed: 21278782
PubMed Central: 4002766
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<author><name sortKey="Wang, Zhong Can" sort="Wang, Zhong Can" uniqKey="Wang Z" first="Zhong-Can" last="Wang">Zhong-Can Wang</name>
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, Nanjing 210029,<country>China</country>
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</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
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<author><name sortKey="Guan, Xiao Hong" sort="Guan, Xiao Hong" uniqKey="Guan X" first="Xiao-Hong" last="Guan">Xiao-Hong Guan</name>
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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>
</TEI>
<pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Acta Pharmacol Sin</journal-id>
<journal-id journal-id-type="iso-abbrev">Acta Pharmacol. Sin</journal-id>
<journal-title-group><journal-title>Acta Pharmacologica Sinica</journal-title>
</journal-title-group>
<issn pub-type="ppub">1671-4083</issn>
<issn pub-type="epub">1745-7254</issn>
<publisher><publisher-name>Nature Publishing Group</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">21278782</article-id>
<article-id pub-id-type="pmc">4002766</article-id>
<article-id pub-id-type="pii">aps2010209</article-id>
<article-id pub-id-type="doi">10.1038/aps.2010.209</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Generation and characterization of the human neutralizing antibody fragment Fab091 against rabies virus</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Li</surname>
<given-names>Chen</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff3">3</xref>
<xref ref-type="aff" rid="aff4">4</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Zhang</surname>
<given-names>Feng</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff3">3</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Lin</surname>
<given-names>Hong</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wang</surname>
<given-names>Zhong-can</given-names>
</name>
<xref ref-type="aff" rid="aff2">2</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Liu</surname>
<given-names>Xin-jian</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Feng</surname>
<given-names>Zhen-qing</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff3">3</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Zhu</surname>
<given-names>Jin</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff2">2</xref>
<xref ref-type="corresp" rid="caf1">*</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Guan</surname>
<given-names>Xiao-hong</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="corresp" rid="caf2">*</xref>
</contrib>
<aff id="aff1"><label>1</label>
<institution>Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University</institution>
, Nanjing 210029,<country>China</country>
</aff>
<aff id="aff2"><label>2</label>
<institution>Huadong Medical Institute of Biotechniques</institution>
, Nanjing 210002,<country>China</country>
</aff>
<aff id="aff3"><label>3</label>
<institution>Department of Pathology, Nanjing Medical University</institution>
, Nanjing 210029,<country>China</country>
</aff>
<aff id="aff4"><label>4</label>
<institution>Department of Pathology, the Affiliated Hospital of Xuzhou Medical College</institution>
, Xuzhou 221002,<country>China</country>
</aff>
</contrib-group>
<author-notes><corresp id="caf1"><label>*</label>
E-mail <email>zjsimmons@yahoo.com.cn</email>
</corresp>
<corresp id="caf2"><label>*</label>
<email>xhguan@163.com</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub"><month>03</month>
<year>2011</year>
</pub-date>
<pub-date pub-type="epub"><day>31</day>
<month>01</month>
<year>2011</year>
</pub-date>
<volume>32</volume>
<issue>3</issue>
<fpage>329</fpage>
<lpage>337</lpage>
<history><date date-type="received"><day>31</day>
<month>05</month>
<year>2010</year>
</date>
<date date-type="accepted"><day>14</day>
<month>11</month>
<year>2010</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2011 CPS and SIMM</copyright-statement>
<copyright-year>2011</copyright-year>
<copyright-holder>CPS and SIMM</copyright-holder>
</permissions>
<abstract><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>
</abstract>
<kwd-group><kwd>rabies</kwd>
<kwd>Fab engineered antibody</kwd>
<kwd>neutralizing antibodies</kwd>
</kwd-group>
</article-meta>
</front>
</pmc>
<affiliations><list><country><li>République populaire de Chine</li>
</country>
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<name sortKey="Feng, Zhen Qing" sort="Feng, Zhen Qing" uniqKey="Feng Z" first="Zhen-Qing" last="Feng">Zhen-Qing Feng</name>
<name sortKey="Feng, Zhen Qing" sort="Feng, Zhen Qing" uniqKey="Feng Z" first="Zhen-Qing" last="Feng">Zhen-Qing Feng</name>
<name sortKey="Guan, Xiao Hong" sort="Guan, Xiao Hong" uniqKey="Guan X" first="Xiao-Hong" last="Guan">Xiao-Hong Guan</name>
<name sortKey="Li, Chen" sort="Li, Chen" uniqKey="Li C" first="Chen" last="Li">Chen Li</name>
<name sortKey="Li, Chen" sort="Li, Chen" uniqKey="Li C" first="Chen" last="Li">Chen Li</name>
<name sortKey="Lin, Hong" sort="Lin, Hong" uniqKey="Lin H" first="Hong" last="Lin">Hong Lin</name>
<name sortKey="Liu, Xin Jian" sort="Liu, Xin Jian" uniqKey="Liu X" first="Xin-Jian" last="Liu">Xin-Jian Liu</name>
<name sortKey="Wang, Zhong Can" sort="Wang, Zhong Can" uniqKey="Wang Z" first="Zhong-Can" last="Wang">Zhong-Can Wang</name>
<name sortKey="Zhang, Feng" sort="Zhang, Feng" uniqKey="Zhang F" first="Feng" last="Zhang">Feng Zhang</name>
<name sortKey="Zhang, Feng" sort="Zhang, Feng" uniqKey="Zhang F" first="Feng" last="Zhang">Feng Zhang</name>
<name sortKey="Zhu, Jin" sort="Zhu, Jin" uniqKey="Zhu J" first="Jin" last="Zhu">Jin Zhu</name>
<name sortKey="Zhu, Jin" sort="Zhu, Jin" uniqKey="Zhu J" first="Jin" last="Zhu">Jin Zhu</name>
</country>
</tree>
</affiliations>
</record>
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