Polyclonal antibodies for specific detection of tobacco host cell proteins can be efficiently generated following RuBisCO depletion and the removal of endotoxins.
Identifieur interne : 000317 ( Main/Corpus ); précédent : 000316; suivant : 000318Polyclonal antibodies for specific detection of tobacco host cell proteins can be efficiently generated following RuBisCO depletion and the removal of endotoxins.
Auteurs : Zulfaquar Ahmad Arfi ; Stephan Hellwig ; Jürgen Drossard ; Rainer Fischer ; Johannes Felix BuyelSource :
- Biotechnology journal [ 1860-7314 ] ; 2016.
English descriptors
- KwdEn :
- Agrobacterium tumefaciens (metabolism), Animals (MeSH), Antibodies, Monoclonal (metabolism), Antibody Specificity (MeSH), Endotoxins (MeSH), Plant Extracts (chemistry), Plant Extracts (immunology), Plant Extracts (isolation & purification), Plants, Genetically Modified (enzymology), Plants, Genetically Modified (microbiology), Polymyxin B (isolation & purification), Rabbits (MeSH), Ribulose-Bisphosphate Carboxylase (deficiency), Tobacco (genetics), Tobacco (immunology), Tobacco (microbiology).
- MESH :
- chemical , chemistry : Plant Extracts.
- chemical , deficiency : Ribulose-Bisphosphate Carboxylase.
- chemical , immunology : Plant Extracts.
- chemical , isolation & purification : Plant Extracts, Polymyxin B.
- chemical , metabolism : Antibodies, Monoclonal.
- enzymology : Plants, Genetically Modified.
- genetics : Tobacco.
- immunology : Tobacco.
- metabolism : Agrobacterium tumefaciens.
- microbiology : Plants, Genetically Modified, Tobacco.
- Animals, Antibody Specificity, Endotoxins, Rabbits.
Abstract
The production of biopharmaceutical proteins in plants requires efficient downstream processing steps that remove impurities such as host cell proteins (HCPs) and adventitious endotoxins produced by bacteria during transient expression. We therefore strived to develop effective routines for endotoxin removal from plant extracts and the subsequent use of the extracts to generate antibodies detecting a broad set of HCPs. At first, we depleted the superabundant protein ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) for which PEG precipitation achieved the best results, preventing a dominant immune reaction against this protein. We found that a mixture of sera from rabbits immunized with pre-depleted or post-depleted extracts detected more HCPs than the individual sera used alone. We also developed a powerful endotoxin removal procedure using Polymyxin B for extracts from wild type plants or a combination of fiber-flow filtration and EndoTrap Blue for tobacco plants infiltrated with Agrobacterium tumefaciens. The antibodies we generated will be useful for quality and performance assessment in future process development and the methods we present can easily be transferred to other expression systems rendering them useful in the field of plant molecular farming.
DOI: 10.1002/biot.201500271
PubMed: 26632519
Links to Exploration step
pubmed:26632519Le document en format XML
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Jurgen" uniqKey="Drossard J" first="Jürgen" last="Drossard">Jürgen Drossard</name><affiliation><nlm:affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Institute for Molecular Biotechnology, RWTH Aachen University, Aachen, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Fischer, Rainer" sort="Fischer, Rainer" uniqKey="Fischer R" first="Rainer" last="Fischer">Rainer Fischer</name><affiliation><nlm:affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Institute for Molecular Biotechnology, RWTH Aachen University, Aachen, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Buyel, Johannes Felix" sort="Buyel, Johannes Felix" uniqKey="Buyel J" first="Johannes Felix" last="Buyel">Johannes Felix Buyel</name><affiliation><nlm:affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany. johannes.buyel@rwth-aachen.de.</nlm:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2016">2016</date><idno type="RBID">pubmed:26632519</idno><idno type="pmid">26632519</idno><idno type="doi">10.1002/biot.201500271</idno><idno type="wicri:Area/Main/Corpus">000317</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000317</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Polyclonal antibodies for specific detection of tobacco host cell proteins can be efficiently generated following RuBisCO depletion and the removal of endotoxins.</title><author><name sortKey="Arfi, Zulfaquar Ahmad" sort="Arfi, Zulfaquar Ahmad" uniqKey="Arfi Z" first="Zulfaquar Ahmad" last="Arfi">Zulfaquar Ahmad Arfi</name><affiliation><nlm:affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>BSV Bioscience GmbH, Baesweiler, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Hellwig, Stephan" sort="Hellwig, Stephan" uniqKey="Hellwig S" first="Stephan" last="Hellwig">Stephan Hellwig</name><affiliation><nlm:affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Drossard, Jurgen" sort="Drossard, Jurgen" uniqKey="Drossard J" first="Jürgen" last="Drossard">Jürgen Drossard</name><affiliation><nlm:affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Institute for Molecular Biotechnology, RWTH Aachen University, Aachen, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Fischer, Rainer" sort="Fischer, Rainer" uniqKey="Fischer R" first="Rainer" last="Fischer">Rainer Fischer</name><affiliation><nlm:affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Institute for Molecular Biotechnology, RWTH Aachen University, Aachen, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Buyel, Johannes Felix" sort="Buyel, Johannes Felix" uniqKey="Buyel J" first="Johannes Felix" last="Buyel">Johannes Felix Buyel</name><affiliation><nlm:affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany. johannes.buyel@rwth-aachen.de.</nlm:affiliation></affiliation></author></analytic><series><title level="j">Biotechnology journal</title><idno type="eISSN">1860-7314</idno><imprint><date when="2016" type="published">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium tumefaciens (metabolism)</term><term>Animals (MeSH)</term><term>Antibodies, Monoclonal (metabolism)</term><term>Antibody Specificity (MeSH)</term><term>Endotoxins (MeSH)</term><term>Plant Extracts (chemistry)</term><term>Plant Extracts (immunology)</term><term>Plant Extracts (isolation & purification)</term><term>Plants, Genetically Modified (enzymology)</term><term>Plants, Genetically Modified (microbiology)</term><term>Polymyxin B (isolation & purification)</term><term>Rabbits (MeSH)</term><term>Ribulose-Bisphosphate Carboxylase (deficiency)</term><term>Tobacco (genetics)</term><term>Tobacco (immunology)</term><term>Tobacco (microbiology)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Plant Extracts</term></keywords><keywords scheme="MESH" type="chemical" qualifier="deficiency" xml:lang="en"><term>Ribulose-Bisphosphate Carboxylase</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Plant Extracts</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Plant Extracts</term><term>Polymyxin B</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Antibodies, Monoclonal</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Plants, Genetically Modified</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Agrobacterium tumefaciens</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Plants, Genetically Modified</term><term>Tobacco</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Antibody Specificity</term><term>Endotoxins</term><term>Rabbits</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The production of biopharmaceutical proteins in plants requires efficient downstream processing steps that remove impurities such as host cell proteins (HCPs) and adventitious endotoxins produced by bacteria during transient expression. We therefore strived to develop effective routines for endotoxin removal from plant extracts and the subsequent use of the extracts to generate antibodies detecting a broad set of HCPs. At first, we depleted the superabundant protein ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) for which PEG precipitation achieved the best results, preventing a dominant immune reaction against this protein. We found that a mixture of sera from rabbits immunized with pre-depleted or post-depleted extracts detected more HCPs than the individual sera used alone. We also developed a powerful endotoxin removal procedure using Polymyxin B for extracts from wild type plants or a combination of fiber-flow filtration and EndoTrap Blue for tobacco plants infiltrated with Agrobacterium tumefaciens. The antibodies we generated will be useful for quality and performance assessment in future process development and the methods we present can easily be transferred to other expression systems rendering them useful in the field of plant molecular farming. </div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">26632519</PMID><DateCompleted><Year>2016</Year><Month>12</Month><Day>16</Day></DateCompleted><DateRevised><Year>2018</Year><Month>12</Month><Day>02</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1860-7314</ISSN><JournalIssue CitedMedium="Internet"><Volume>11</Volume><Issue>4</Issue><PubDate><Year>2016</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Biotechnology journal</Title><ISOAbbreviation>Biotechnol J</ISOAbbreviation></Journal><ArticleTitle>Polyclonal antibodies for specific detection of tobacco host cell proteins can be efficiently generated following RuBisCO depletion and the removal of endotoxins.</ArticleTitle><Pagination><MedlinePgn>507-18</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1002/biot.201500271</ELocationID><Abstract><AbstractText>The production of biopharmaceutical proteins in plants requires efficient downstream processing steps that remove impurities such as host cell proteins (HCPs) and adventitious endotoxins produced by bacteria during transient expression. We therefore strived to develop effective routines for endotoxin removal from plant extracts and the subsequent use of the extracts to generate antibodies detecting a broad set of HCPs. At first, we depleted the superabundant protein ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) for which PEG precipitation achieved the best results, preventing a dominant immune reaction against this protein. We found that a mixture of sera from rabbits immunized with pre-depleted or post-depleted extracts detected more HCPs than the individual sera used alone. We also developed a powerful endotoxin removal procedure using Polymyxin B for extracts from wild type plants or a combination of fiber-flow filtration and EndoTrap Blue for tobacco plants infiltrated with Agrobacterium tumefaciens. The antibodies we generated will be useful for quality and performance assessment in future process development and the methods we present can easily be transferred to other expression systems rendering them useful in the field of plant molecular farming. </AbstractText><CopyrightInformation>Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Arfi</LastName><ForeName>Zulfaquar Ahmad</ForeName><Initials>ZA</Initials><AffiliationInfo><Affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>BSV Bioscience GmbH, Baesweiler, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hellwig</LastName><ForeName>Stephan</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Drossard</LastName><ForeName>Jürgen</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Institute for Molecular Biotechnology, RWTH Aachen University, Aachen, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Fischer</LastName><ForeName>Rainer</ForeName><Initials>R</Initials><AffiliationInfo><Affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Institute for Molecular Biotechnology, RWTH Aachen University, Aachen, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Buyel</LastName><ForeName>Johannes Felix</ForeName><Initials>JF</Initials><AffiliationInfo><Affiliation>Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany. johannes.buyel@rwth-aachen.de.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2016</Year><Month>01</Month><Day>14</Day></ArticleDate></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>Biotechnol J</MedlineTA><NlmUniqueID>101265833</NlmUniqueID><ISSNLinking>1860-6768</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000911">Antibodies, Monoclonal</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D004731">Endotoxins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010936">Plant Extracts</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 4.1.1.39</RegistryNumber><NameOfSubstance UI="D012273">Ribulose-Bisphosphate Carboxylase</NameOfSubstance></Chemical><Chemical><RegistryNumber>J2VZ07J96K</RegistryNumber><NameOfSubstance UI="D011112">Polymyxin B</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D016960" MajorTopicYN="N">Agrobacterium tumefaciens</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000911" MajorTopicYN="N">Antibodies, Monoclonal</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D000918" MajorTopicYN="N">Antibody Specificity</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004731" MajorTopicYN="N">Endotoxins</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010936" MajorTopicYN="N">Plant Extracts</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D030821" MajorTopicYN="N">Plants, Genetically Modified</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="N">enzymology</QualifierName><QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011112" MajorTopicYN="N">Polymyxin B</DescriptorName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011817" MajorTopicYN="N">Rabbits</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012273" MajorTopicYN="N">Ribulose-Bisphosphate Carboxylase</DescriptorName><QualifierName UI="Q000172" MajorTopicYN="Y">deficiency</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014026" MajorTopicYN="N">Tobacco</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName><QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Endotoxin removal</Keyword><Keyword MajorTopicYN="N">Molecular farming</Keyword><Keyword MajorTopicYN="N">Plant-derived biopharmaceutical proteins</Keyword><Keyword MajorTopicYN="N">polyclonal anti-host cell protein antibodies</Keyword><Keyword MajorTopicYN="N">quality control</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2015</Year><Month>05</Month><Day>04</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2015</Year><Month>08</Month><Day>21</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2015</Year><Month>12</Month><Day>02</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2015</Year><Month>12</Month><Day>4</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2015</Year><Month>12</Month><Day>4</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2016</Year><Month>12</Month><Day>17</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">26632519</ArticleId><ArticleId IdType="doi">10.1002/biot.201500271</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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