[Expression and renaturation of a novel human single-chain Fv antibody against SARS-CoV].
Identifieur interne : 001259 ( Ncbi/Checkpoint ); précédent : 001258; suivant : 001260[Expression and renaturation of a novel human single-chain Fv antibody against SARS-CoV].
Auteurs : Jin-Zhu Duan [République populaire de Chine] ; Cai Qi ; Wei Han ; Zhan-Hui Wang ; Gang Jin ; Xi-Yun YanSource :
- Sheng wu gong cheng xue bao = Chinese journal of biotechnology [ 1000-3061 ] ; 2005.
Descripteurs français
- KwdFr :
- Anticorps antiviraux (immunologie), Anticorps monoclonaux (biosynthèse), Anticorps monoclonaux (génétique), Corps d'inclusion (génétique), Corps d'inclusion (immunologie), Escherichia coli (génétique), Escherichia coli (métabolisme), Fragments d'immunoglobuline (biosynthèse), Fragments d'immunoglobuline (génétique), Fragments d'immunoglobuline (immunologie), Humains, Protéines recombinantes (), Protéines recombinantes (biosynthèse), Protéines recombinantes (immunologie), Renaturation des protéines, Région variable d'immunoglobuline (biosynthèse), Région variable d'immunoglobuline (génétique), Région variable d'immunoglobuline (immunologie), Virus du SRAS (immunologie).
- MESH :
- biosynthèse : Anticorps monoclonaux, Fragments d'immunoglobuline, Protéines recombinantes, Région variable d'immunoglobuline.
- génétique : Anticorps monoclonaux, Corps d'inclusion, Escherichia coli, Fragments d'immunoglobuline, Région variable d'immunoglobuline.
- immunologie : Anticorps antiviraux, Corps d'inclusion, Fragments d'immunoglobuline, Protéines recombinantes, Région variable d'immunoglobuline, Virus du SRAS.
- métabolisme : Escherichia coli.
- Humains, Protéines recombinantes, Renaturation des protéines.
English descriptors
- KwdEn :
- Antibodies, Monoclonal (biosynthesis), Antibodies, Monoclonal (genetics), Antibodies, Viral (immunology), Escherichia coli (genetics), Escherichia coli (metabolism), Humans, Immunoglobulin Fragments (biosynthesis), Immunoglobulin Fragments (genetics), Immunoglobulin Fragments (immunology), Immunoglobulin Variable Region (biosynthesis), Immunoglobulin Variable Region (genetics), Immunoglobulin Variable Region (immunology), Inclusion Bodies (genetics), Inclusion Bodies (immunology), Protein Renaturation, Recombinant Proteins (biosynthesis), Recombinant Proteins (chemistry), Recombinant Proteins (immunology), SARS Virus (immunology).
- MESH :
- chemical , biosynthesis : Antibodies, Monoclonal, Immunoglobulin Fragments, Immunoglobulin Variable Region, Recombinant Proteins.
- chemical , chemistry : Recombinant Proteins.
- chemical , genetics : Antibodies, Monoclonal, Immunoglobulin Fragments, Immunoglobulin Variable Region.
- chemical , immunology : Antibodies, Viral, Immunoglobulin Fragments, Immunoglobulin Variable Region, Recombinant Proteins.
- genetics : Escherichia coli, Inclusion Bodies.
- immunology : Inclusion Bodies, SARS Virus.
- metabolism : Escherichia coli.
- Humans, Protein Renaturation.
Abstract
A novel human ScFv H12 against SARS-CoV has been selected from a SARS immune library. In order to produce a large amount of ScFv H12, pET28a-H12 expression vector was constructed and ScFv H12 was expressed at yield about 30% of total proteins in E. coli . Here two different refolding procedures were used to refold ScFv H12 from inclusion body: gel filtration chromatography and dilution. The results showed that ScFv H12 could be efficiently refolded by both procedures. However, the refolding via gel filtration was 1.5 time more effective than that of dilution. The affinity of ScFv H12 to SARS-CoV virion was detected as Kd = 73.5 nmol/mL.
PubMed: 16285506
Affiliations:
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pubmed:16285506Le document en format XML
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sortKey="Qi, Cai" sort="Qi, Cai" uniqKey="Qi C" first="Cai" last="Qi">Cai Qi</name></author><author><name sortKey="Han, Wei" sort="Han, Wei" uniqKey="Han W" first="Wei" last="Han">Wei Han</name></author><author><name sortKey="Wang, Zhan Hui" sort="Wang, Zhan Hui" uniqKey="Wang Z" first="Zhan-Hui" last="Wang">Zhan-Hui Wang</name></author><author><name sortKey="Jin, Gang" sort="Jin, Gang" uniqKey="Jin G" first="Gang" last="Jin">Gang Jin</name></author><author><name sortKey="Yan, Xi Yun" sort="Yan, Xi Yun" uniqKey="Yan X" first="Xi-Yun" last="Yan">Xi-Yun Yan</name></author></analytic><series><title level="j">Sheng wu gong cheng xue bao = Chinese journal of biotechnology</title><idno type="ISSN">1000-3061</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Monoclonal (biosynthesis)</term><term>Antibodies, Monoclonal 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protéines</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A novel human ScFv H12 against SARS-CoV has been selected from a SARS immune library. In order to produce a large amount of ScFv H12, pET28a-H12 expression vector was constructed and ScFv H12 was expressed at yield about 30% of total proteins in E. coli . Here two different refolding procedures were used to refold ScFv H12 from inclusion body: gel filtration chromatography and dilution. The results showed that ScFv H12 could be efficiently refolded by both procedures. However, the refolding via gel filtration was 1.5 time more effective than that of dilution. 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