Sequential affinity purification of peroxidase tagged bispecific anti-SARS-CoV antibodies on phenylboronic acid agarose
Identifieur interne : 003475 ( Main/Exploration ); précédent : 003474; suivant : 003476Sequential affinity purification of peroxidase tagged bispecific anti-SARS-CoV antibodies on phenylboronic acid agarose
Auteurs : Pravin K. Bhatnagar [Canada] ; Dipankar Das [Canada] ; Mavanur R. Suresh [Canada]Source :
- Journal of chromatography. B [ 1570-0232 ] ; 2008.
Descripteurs français
- KwdFr :
- Affinité des anticorps, Agarose (), Agarose (analogues et dérivés), Animaux, Anticorps antiviraux (isolement et purification), Anticorps bispécifiques (isolement et purification), Anticorps monoclonaux (immunologie), Anticorps monoclonaux (isolement et purification), Chromatographie d'affinité (), Horseradish peroxidase (), Hybridomes (immunologie), Rats, Sites de fixation des anticorps, Techniques immunoenzymatiques, Test ELISA (), Virus du SRAS (immunologie).
- MESH :
- analogues et dérivés : Agarose.
- immunologie : Anticorps monoclonaux, Hybridomes, Virus du SRAS.
- isolement et purification : Anticorps antiviraux, Anticorps bispécifiques, Anticorps monoclonaux.
- Pascal (Inist)
- Affinité, Affinité des anticorps, Agarose, Animaux, Chromatographie d'affinité, Horseradish peroxidase, Purification, Peroxidase, Antiviral, Rats, Sites de fixation des anticorps, Techniques immunoenzymatiques, Test ELISA, Virus syndrome respiratoire aigu sévère, Anticorps bispécifique, Agarose, Anticorps monoclonal, Boronique acide, Borane(aminophényl dihydroxy).
English descriptors
- KwdEn :
- Affinity, Agarose, Animals, Antibodies, Bispecific (isolation & purification), Antibodies, Monoclonal (immunology), Antibodies, Monoclonal (isolation & purification), Antibodies, Viral (isolation & purification), Antibody Affinity, Antiviral, Binding Sites, Antibody, Bispecific antibody, Chromatography, Affinity (methods), Enzyme-Linked Immunosorbent Assay (methods), Horseradish Peroxidase (chemistry), Hybridomas (immunology), Immunoenzyme Techniques, Monoclonal antibody, Peroxidase, Purification, Rats, SARS Virus (immunology), Sepharose (analogs & derivatives), Sepharose (chemistry), Severe acute respiratory syndrome virus.
- MESH :
- chemical , analogs & derivatives : Sepharose.
- chemical , chemistry : Horseradish Peroxidase, Sepharose.
- chemical , immunology : Antibodies, Monoclonal.
- chemical , isolation & purification : Antibodies, Bispecific, Antibodies, Monoclonal, Antibodies, Viral.
- immunology : Hybridomas, SARS Virus.
- methods : Chromatography, Affinity, Enzyme-Linked Immunosorbent Assay.
- Animals, Antibody Affinity, Binding Sites, Antibody, Immunoenzyme Techniques, Rats.
Abstract
Hybrid hybridomas (quadromas) are derived by fusing at least two hybridomas, each producing a different antibody of predefined specificity. The resulting cell secretes not only the immunoglobulins of both parents but also hybrid molecules manifesting the binding characteristics of the individual fusion partners. Purification of the desired bispecific immunoprobe with high specific activity from a mixture of bispecific and monospecific monoclonal antibodies requires special strategies. Using a dual, sequential affinity chromatography (Protein-G chromatography followed by m-aminophenyleboronic acid agarose column), we have purified bispecific monoclonal antibodies (BsMAb) as a preformed HRPO (Horseradish Peroxidase) complex (BsMAb-HRPO). The quadroma culture supernatant was initially processed on a Protein-G column to isolate all the species of immunoglobulins. This pre-enriched fraction was subsequently passed through the aminophenyleboronic acid column super saturated with HRPO. The column matrix has the ability to bind to proteins such as HRPO with vicinal diols. The enzyme loaded column captures the desired bispecific anti-SARS-CoV x anti-HRPO species with the elimination of the monospecific anti-SARS-CoV MAb to result in a high specific activity diagnostic probe. The presence of anti-HRPO MAb is an acceptable impurity as it will not bind to the target SARS-CoV NP antigen and will get washed out during the ELISA procedure.
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream PascalFrancis, to step Corpus: 000306
- to stream PascalFrancis, to step Curation: 000682
- to stream PascalFrancis, to step Checkpoint: 000255
- to stream Main, to step Merge: 003581
- to stream PubMed, to step Corpus: 001C13
- to stream PubMed, to step Curation: 001C13
- to stream PubMed, to step Checkpoint: 001A28
- to stream Ncbi, to step Merge: 001B78
- to stream Ncbi, to step Curation: 001B78
- to stream Ncbi, to step Checkpoint: 001B78
- to stream Main, to step Merge: 003099
- to stream Main, to step Curation: 003475
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en" level="a">Sequential affinity purification of peroxidase tagged bispecific anti-SARS-CoV antibodies on phenylboronic acid agarose</title><author><name sortKey="Bhatnagar, Pravin K" sort="Bhatnagar, Pravin K" uniqKey="Bhatnagar P" first="Pravin K." last="Bhatnagar">Pravin K. Bhatnagar</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta</s1><s2>Edmonton, Alberta, T6G 2N8</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Edmonton, Alberta, T6G 2N8</wicri:noRegion></affiliation></author><author><name sortKey="Das, Dipankar" sort="Das, Dipankar" uniqKey="Das D" first="Dipankar" last="Das">Dipankar Das</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta</s1><s2>Edmonton, Alberta, T6G 2N8</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Edmonton, Alberta, T6G 2N8</wicri:noRegion></affiliation></author><author><name sortKey="Suresh, Mavanur R" sort="Suresh, Mavanur R" uniqKey="Suresh M" first="Mavanur R." last="Suresh">Mavanur R. Suresh</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta</s1><s2>Edmonton, Alberta, T6G 2N8</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Edmonton, Alberta, T6G 2N8</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">08-0161046</idno><date when="2008">2008</date><idno type="stanalyst">PASCAL 08-0161046 INIST</idno><idno type="RBID">Pascal:08-0161046</idno><idno type="wicri:Area/PascalFrancis/Corpus">000306</idno><idno type="wicri:Area/PascalFrancis/Curation">000682</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000255</idno><idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000255</idno><idno type="wicri:doubleKey">1570-0232:2008:Bhatnagar P:sequential:affinity:purification</idno><idno type="wicri:Area/Main/Merge">003581</idno><idno type="wicri:source">PubMed</idno><idno type="RBID">pubmed:18258500</idno><idno type="wicri:Area/PubMed/Corpus">001C13</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001C13</idno><idno type="wicri:Area/PubMed/Curation">001C13</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001C13</idno><idno type="wicri:Area/PubMed/Checkpoint">001A28</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001A28</idno><idno type="wicri:Area/Ncbi/Merge">001B78</idno><idno type="wicri:Area/Ncbi/Curation">001B78</idno><idno type="wicri:Area/Ncbi/Checkpoint">001B78</idno><idno type="wicri:doubleKey">1570-0232:2008:Bhatnagar P:sequential:affinity:purification</idno><idno type="wicri:Area/Main/Merge">003099</idno><idno type="wicri:Area/Main/Curation">003475</idno><idno type="wicri:Area/Main/Exploration">003475</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Sequential affinity purification of peroxidase tagged bispecific anti-SARS-CoV antibodies on phenylboronic acid agarose</title><author><name sortKey="Bhatnagar, Pravin K" sort="Bhatnagar, Pravin K" uniqKey="Bhatnagar P" first="Pravin K." last="Bhatnagar">Pravin K. Bhatnagar</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta</s1><s2>Edmonton, Alberta, T6G 2N8</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Edmonton, Alberta, T6G 2N8</wicri:noRegion></affiliation></author><author><name sortKey="Das, Dipankar" sort="Das, Dipankar" uniqKey="Das D" first="Dipankar" last="Das">Dipankar Das</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta</s1><s2>Edmonton, Alberta, T6G 2N8</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Edmonton, Alberta, T6G 2N8</wicri:noRegion></affiliation></author><author><name sortKey="Suresh, Mavanur R" sort="Suresh, Mavanur R" uniqKey="Suresh M" first="Mavanur R." last="Suresh">Mavanur R. Suresh</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta</s1><s2>Edmonton, Alberta, T6G 2N8</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Edmonton, Alberta, T6G 2N8</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">Journal of chromatography. B</title><title level="j" type="abbreviated">J. chromatogr., B</title><idno type="ISSN">1570-0232</idno><imprint><date when="2008">2008</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of chromatography. B</title><title level="j" type="abbreviated">J. chromatogr., B</title><idno type="ISSN">1570-0232</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Affinity</term><term>Agarose</term><term>Animals</term><term>Antibodies, Bispecific (isolation & purification)</term><term>Antibodies, Monoclonal (immunology)</term><term>Antibodies, Monoclonal (isolation & purification)</term><term>Antibodies, Viral (isolation & purification)</term><term>Antibody Affinity</term><term>Antiviral</term><term>Binding Sites, Antibody</term><term>Bispecific antibody</term><term>Chromatography, Affinity (methods)</term><term>Enzyme-Linked Immunosorbent Assay (methods)</term><term>Horseradish Peroxidase (chemistry)</term><term>Hybridomas (immunology)</term><term>Immunoenzyme Techniques</term><term>Monoclonal antibody</term><term>Peroxidase</term><term>Purification</term><term>Rats</term><term>SARS Virus (immunology)</term><term>Sepharose (analogs & derivatives)</term><term>Sepharose (chemistry)</term><term>Severe acute respiratory syndrome virus</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Affinité des anticorps</term><term>Agarose ()</term><term>Agarose (analogues et dérivés)</term><term>Animaux</term><term>Anticorps antiviraux (isolement et purification)</term><term>Anticorps bispécifiques (isolement et purification)</term><term>Anticorps monoclonaux (immunologie)</term><term>Anticorps monoclonaux (isolement et purification)</term><term>Chromatographie d'affinité ()</term><term>Horseradish peroxidase ()</term><term>Hybridomes (immunologie)</term><term>Rats</term><term>Sites de fixation des anticorps</term><term>Techniques immunoenzymatiques</term><term>Test ELISA ()</term><term>Virus du SRAS (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analogs & derivatives" xml:lang="en"><term>Sepharose</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Horseradish Peroxidase</term><term>Sepharose</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Monoclonal</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Antibodies, Bispecific</term><term>Antibodies, Monoclonal</term><term>Antibodies, Viral</term></keywords><keywords scheme="MESH" qualifier="analogues et dérivés" xml:lang="fr"><term>Agarose</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps monoclonaux</term><term>Hybridomes</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Hybridomas</term><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Anticorps antiviraux</term><term>Anticorps bispécifiques</term><term>Anticorps monoclonaux</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Chromatography, Affinity</term><term>Enzyme-Linked Immunosorbent Assay</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Antibody Affinity</term><term>Binding Sites, Antibody</term><term>Immunoenzyme Techniques</term><term>Rats</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Affinité</term><term>Affinité des anticorps</term><term>Agarose</term><term>Animaux</term><term>Chromatographie d'affinité</term><term>Horseradish peroxidase</term><term>Purification</term><term>Peroxidase</term><term>Antiviral</term><term>Rats</term><term>Sites de fixation des anticorps</term><term>Techniques immunoenzymatiques</term><term>Test ELISA</term><term>Virus syndrome respiratoire aigu sévère</term><term>Anticorps bispécifique</term><term>Agarose</term><term>Anticorps monoclonal</term><term>Boronique acide</term><term>Borane(aminophényl dihydroxy)</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Hybrid hybridomas (quadromas) are derived by fusing at least two hybridomas, each producing a different antibody of predefined specificity. The resulting cell secretes not only the immunoglobulins of both parents but also hybrid molecules manifesting the binding characteristics of the individual fusion partners. Purification of the desired bispecific immunoprobe with high specific activity from a mixture of bispecific and monospecific monoclonal antibodies requires special strategies. Using a dual, sequential affinity chromatography (Protein-G chromatography followed by m-aminophenyleboronic acid agarose column), we have purified bispecific monoclonal antibodies (BsMAb) as a preformed HRPO (Horseradish Peroxidase) complex (BsMAb-HRPO). The quadroma culture supernatant was initially processed on a Protein-G column to isolate all the species of immunoglobulins. This pre-enriched fraction was subsequently passed through the aminophenyleboronic acid column super saturated with HRPO. The column matrix has the ability to bind to proteins such as HRPO with vicinal diols. The enzyme loaded column captures the desired bispecific anti-SARS-CoV x anti-HRPO species with the elimination of the monospecific anti-SARS-CoV MAb to result in a high specific activity diagnostic probe. The presence of anti-HRPO MAb is an acceptable impurity as it will not bind to the target SARS-CoV NP antigen and will get washed out during the ELISA procedure.</div></front></TEI><affiliations><list><country><li>Canada</li></country></list><tree><country name="Canada"><noRegion><name sortKey="Bhatnagar, Pravin K" sort="Bhatnagar, Pravin K" uniqKey="Bhatnagar P" first="Pravin K." last="Bhatnagar">Pravin K. Bhatnagar</name></noRegion><name sortKey="Das, Dipankar" sort="Das, Dipankar" uniqKey="Das D" first="Dipankar" last="Das">Dipankar Das</name><name sortKey="Suresh, Mavanur R" sort="Suresh, Mavanur R" uniqKey="Suresh M" first="Mavanur R." last="Suresh">Mavanur R. Suresh</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/SrasV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 003475 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 003475 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= SrasV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= Pascal:08-0161046
|texte= Sequential affinity purification of peroxidase tagged bispecific anti-SARS-CoV antibodies on phenylboronic acid agarose
}}
| This area was generated with Dilib version V0.6.33. |  |