SrasV1, Main, Exploration, bibRecord, 000D95

Optimization of the Production Process and Characterization of the Yeast-Expressed SARS-CoV Recombinant Receptor-Binding Domain (RBD219-N1), a SARS Vaccine Candidate.

Identifieur interne : 000D95 ( Main/Exploration ); précédent : 000D94; suivant : 000D96

Optimization of the Production Process and Characterization of the Yeast-Expressed SARS-CoV Recombinant Receptor-Binding Domain (RBD219-N1), a SARS Vaccine Candidate.

Auteurs : Wen-Hsiang Chen [États-Unis] ; Shivali M. Chag [États-Unis] ; Mohan V. Poongavanam [États-Unis] ; Amadeo B. Biter [États-Unis] ; Ebe A. Ewere [États-Unis] ; Wanderson Rezende [États-Unis] ; Christopher A. Seid [États-Unis] ; Elissa M. Hudspeth [États-Unis] ; Jeroen Pollet [États-Unis] ; C Patrick Mcatee [États-Unis] ; Ulrich Strych [États-Unis] ; Maria Elena Bottazzi [États-Unis] ; Peter J. Hotez [États-Unis]

Source :

Journal of pharmaceutical sciences [ 1520-6017 ] ; 2017.

RBID : pubmed:28456726

Descripteurs français

English descriptors

KwdEn :
- Cloning, Molecular (methods), Fermentation, Humans, Industrial Microbiology (methods), Pichia (genetics), Protein Domains, Recombinant Proteins (chemistry), Recombinant Proteins (genetics), Recombinant Proteins (isolation & purification), SARS Virus (chemistry), SARS Virus (genetics), Severe Acute Respiratory Syndrome (prevention & control), Severe Acute Respiratory Syndrome (virology), Spike Glycoprotein, Coronavirus (chemistry), Spike Glycoprotein, Coronavirus (genetics), Spike Glycoprotein, Coronavirus (isolation & purification), Vaccines, Synthetic (chemistry), Vaccines, Synthetic (genetics), Vaccines, Synthetic (isolation & purification), Viral Vaccines (chemistry), Viral Vaccines (genetics), Viral Vaccines (isolation & purification).
MESH :
- chemical , chemistry : Recombinant Proteins, Spike Glycoprotein, Coronavirus, Vaccines, Synthetic, Viral Vaccines.
- chemistry : SARS Virus.
- genetics : Pichia, Recombinant Proteins, SARS Virus, Spike Glycoprotein, Coronavirus, Vaccines, Synthetic, Viral Vaccines.
- chemical , isolation & purification : Recombinant Proteins, Spike Glycoprotein, Coronavirus, Vaccines, Synthetic, Viral Vaccines.
- methods : Cloning, Molecular, Industrial Microbiology.
- prevention & control : Severe Acute Respiratory Syndrome.
- virology : Severe Acute Respiratory Syndrome.
- Fermentation, Humans, Protein Domains.

Abstract

From 2002 to 2003, a global pandemic of severe acute respiratory syndrome (SARS) spread to 5 continents and caused 8000 respiratory infections and 800 deaths. To ameliorate the effects of future outbreaks as well as to prepare for biodefense, a process for the production of a recombinant protein vaccine candidate is under development. Previously, we reported the 5 L scale expression and purification of a promising recombinant SARS vaccine candidate, RBD219-N1, the 218-amino acid residue receptor-binding domain (RBD) of SARS coronavirus expressed in yeast-Pichia pastoris X-33. When adjuvanted with aluminum hydroxide, this protein elicited high neutralizing antibody titers and high RBD-specific antibody titers. However, the yield of RBD219-N1 (60 mg RBD219-N1 per liter of fermentation supernatant; 60 mg/L FS) still required improvement to reach our target of >100 mg/L FS. In this study, we optimized the 10 L scale production process and increased the fermentation yield 6- to 7-fold to 400 mg/L FS with purification recovery >50%. A panel of characterization tests indicated that the process is reproducible and that the purified, tag-free RBD219-N1 protein has high purity and a well-defined structure and is therefore a suitable candidate for production under current Good Manufacturing Practice and future phase-1 clinical trials.

DOI: 10.1016/j.xphs.2017.04.037
PubMed: 28456726

Affiliations:

Le document en format XML

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<sourceDesc><biblStruct><analytic><title xml:lang="en">Optimization of the Production Process and Characterization of the Yeast-Expressed SARS-CoV Recombinant Receptor-Binding Domain (RBD219-N1), a SARS Vaccine Candidate.</title>
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<country xml:lang="fr">États-Unis</country>
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</affiliation>
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<author><name sortKey="Bottazzi, Maria Elena" sort="Bottazzi, Maria Elena" uniqKey="Bottazzi M" first="Maria Elena" last="Bottazzi">Maria Elena Bottazzi</name>
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<term>Fermentation</term>
<term>Humans</term>
<term>Industrial Microbiology (methods)</term>
<term>Pichia (genetics)</term>
<term>Protein Domains</term>
<term>Recombinant Proteins (chemistry)</term>
<term>Recombinant Proteins (genetics)</term>
<term>Recombinant Proteins (isolation & purification)</term>
<term>SARS Virus (chemistry)</term>
<term>SARS Virus (genetics)</term>
<term>Severe Acute Respiratory Syndrome (prevention & control)</term>
<term>Severe Acute Respiratory Syndrome (virology)</term>
<term>Spike Glycoprotein, Coronavirus (chemistry)</term>
<term>Spike Glycoprotein, Coronavirus (genetics)</term>
<term>Spike Glycoprotein, Coronavirus (isolation & purification)</term>
<term>Vaccines, Synthetic (chemistry)</term>
<term>Vaccines, Synthetic (genetics)</term>
<term>Vaccines, Synthetic (isolation & purification)</term>
<term>Viral Vaccines (chemistry)</term>
<term>Viral Vaccines (genetics)</term>
<term>Viral Vaccines (isolation & purification)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Clonage moléculaire ()</term>
<term>Domaines protéiques</term>
<term>Fermentation</term>
<term>Glycoprotéine de spicule des coronavirus ()</term>
<term>Glycoprotéine de spicule des coronavirus (génétique)</term>
<term>Glycoprotéine de spicule des coronavirus (isolement et purification)</term>
<term>Humains</term>
<term>Microbiologie industrielle ()</term>
<term>Pichia (génétique)</term>
<term>Protéines recombinantes ()</term>
<term>Protéines recombinantes (génétique)</term>
<term>Protéines recombinantes (isolement et purification)</term>
<term>Syndrome respiratoire aigu sévère ()</term>
<term>Syndrome respiratoire aigu sévère (virologie)</term>
<term>Vaccins antiviraux ()</term>
<term>Vaccins antiviraux (génétique)</term>
<term>Vaccins antiviraux (isolement et purification)</term>
<term>Vaccins synthétiques ()</term>
<term>Vaccins synthétiques (génétique)</term>
<term>Vaccins synthétiques (isolement et purification)</term>
<term>Virus du SRAS ()</term>
<term>Virus du SRAS (génétique)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Recombinant Proteins</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Vaccines, Synthetic</term>
<term>Viral Vaccines</term>
</keywords>
<keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Pichia</term>
<term>Recombinant Proteins</term>
<term>SARS Virus</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Vaccines, Synthetic</term>
<term>Viral Vaccines</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Glycoprotéine de spicule des coronavirus</term>
<term>Pichia</term>
<term>Protéines recombinantes</term>
<term>Vaccins antiviraux</term>
<term>Vaccins synthétiques</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Recombinant Proteins</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Vaccines, Synthetic</term>
<term>Viral Vaccines</term>
</keywords>
<keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Glycoprotéine de spicule des coronavirus</term>
<term>Protéines recombinantes</term>
<term>Vaccins antiviraux</term>
<term>Vaccins synthétiques</term>
</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Cloning, Molecular</term>
<term>Industrial Microbiology</term>
</keywords>
<keywords scheme="MESH" qualifier="prevention & control" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Fermentation</term>
<term>Humans</term>
<term>Protein Domains</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Clonage moléculaire</term>
<term>Domaines protéiques</term>
<term>Fermentation</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Humains</term>
<term>Microbiologie industrielle</term>
<term>Protéines recombinantes</term>
<term>Syndrome respiratoire aigu sévère</term>
<term>Vaccins antiviraux</term>
<term>Vaccins synthétiques</term>
<term>Virus du SRAS</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">From 2002 to 2003, a global pandemic of severe acute respiratory syndrome (SARS) spread to 5 continents and caused 8000 respiratory infections and 800 deaths. To ameliorate the effects of future outbreaks as well as to prepare for biodefense, a process for the production of a recombinant protein vaccine candidate is under development. Previously, we reported the 5 L scale expression and purification of a promising recombinant SARS vaccine candidate, RBD219-N1, the 218-amino acid residue receptor-binding domain (RBD) of SARS coronavirus expressed in yeast-Pichia pastoris X-33. When adjuvanted with aluminum hydroxide, this protein elicited high neutralizing antibody titers and high RBD-specific antibody titers. However, the yield of RBD219-N1 (60 mg RBD219-N1 per liter of fermentation supernatant; 60 mg/L FS) still required improvement to reach our target of >100 mg/L FS. In this study, we optimized the 10 L scale production process and increased the fermentation yield 6- to 7-fold to 400 mg/L FS with purification recovery >50%. A panel of characterization tests indicated that the process is reproducible and that the purified, tag-free RBD219-N1 protein has high purity and a well-defined structure and is therefore a suitable candidate for production under current Good Manufacturing Practice and future phase-1 clinical trials.</div>
</front>
</TEI>
<affiliations><list><country><li>États-Unis</li>
</country>
<region><li>Texas</li>
</region>
</list>
<tree><country name="États-Unis"><region name="Texas"><name sortKey="Chen, Wen Hsiang" sort="Chen, Wen Hsiang" uniqKey="Chen W" first="Wen-Hsiang" last="Chen">Wen-Hsiang Chen</name>
</region>
<name sortKey="Biter, Amadeo B" sort="Biter, Amadeo B" uniqKey="Biter A" first="Amadeo B" last="Biter">Amadeo B. Biter</name>
<name sortKey="Bottazzi, Maria Elena" sort="Bottazzi, Maria Elena" uniqKey="Bottazzi M" first="Maria Elena" last="Bottazzi">Maria Elena Bottazzi</name>
<name sortKey="Chag, Shivali M" sort="Chag, Shivali M" uniqKey="Chag S" first="Shivali M" last="Chag">Shivali M. Chag</name>
<name sortKey="Ewere, Ebe A" sort="Ewere, Ebe A" uniqKey="Ewere E" first="Ebe A" last="Ewere">Ebe A. Ewere</name>
<name sortKey="Hotez, Peter J" sort="Hotez, Peter J" uniqKey="Hotez P" first="Peter J" last="Hotez">Peter J. Hotez</name>
<name sortKey="Hudspeth, Elissa M" sort="Hudspeth, Elissa M" uniqKey="Hudspeth E" first="Elissa M" last="Hudspeth">Elissa M. Hudspeth</name>
<name sortKey="Mcatee, C Patrick" sort="Mcatee, C Patrick" uniqKey="Mcatee C" first="C Patrick" last="Mcatee">C Patrick Mcatee</name>
<name sortKey="Pollet, Jeroen" sort="Pollet, Jeroen" uniqKey="Pollet J" first="Jeroen" last="Pollet">Jeroen Pollet</name>
<name sortKey="Poongavanam, Mohan V" sort="Poongavanam, Mohan V" uniqKey="Poongavanam M" first="Mohan V" last="Poongavanam">Mohan V. Poongavanam</name>
<name sortKey="Rezende, Wanderson" sort="Rezende, Wanderson" uniqKey="Rezende W" first="Wanderson" last="Rezende">Wanderson Rezende</name>
<name sortKey="Seid, Christopher A" sort="Seid, Christopher A" uniqKey="Seid C" first="Christopher A" last="Seid">Christopher A. Seid</name>
<name sortKey="Strych, Ulrich" sort="Strych, Ulrich" uniqKey="Strych U" first="Ulrich" last="Strych">Ulrich Strych</name>
</country>
</tree>
</affiliations>
</record>

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Optimization of the Production Process and Characterization of the Yeast-Expressed SARS-CoV Recombinant Receptor-Binding Domain (RBD219-N1), a SARS Vaccine Candidate.

Optimization of the Production Process and Characterization of the Yeast-Expressed SARS-CoV Recombinant Receptor-Binding Domain (RBD219-N1), a SARS Vaccine Candidate.

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