Integrated microreactor for enzymatic reaction automation: An easy step toward the quality control of monoclonal antibodies.
Identifieur interne : 000268 ( France/Analysis ); précédent : 000267; suivant : 000269Integrated microreactor for enzymatic reaction automation: An easy step toward the quality control of monoclonal antibodies.
Auteurs : Yoann Ladner [France] ; Silvia Mas [France] ; Gaelle Coussot [France] ; Killian Bartley [France] ; Jérôme Montels [France] ; Jacques Morel [France] ; Catherine Perrin [France]Source :
- Journal of chromatography. A [ 1873-3778 ] ; 2017.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , analysis : Antibodies, Monoclonal, Humanized, Trastuzumab.
- chemical : Antibodies, Monoclonal.
- instrumentation : Chemistry, Pharmaceutical.
- methods : Enzyme Assays.
- Automation, Electrophoresis, Capillary, Hydrogen-Ion Concentration, Osmolar Concentration, Proteolysis, Quality Control.
- mix :
- Antibodies, Automation, Capillary, Capillary electrophoresis, Chemistry, Electrophoresis, Enzymatic microreactor, Enzyme Assays, Humanized, Hydrogen-Ion Concentration, In-line digestion, Monoclonal, Monoclonal antibody, Osmolar Concentration, Pharmaceutical, Proteolysis, Proteolysis product separation, Quality control, Trastuzumab, Quality Control.
Abstract
The main purpose of the present work is to provide a fully integrated miniaturized electrophoretic methodology in order to facilitate the quality control of monoclonal antibodies (mAbs). This methodology called D-PES, which stands for Diffusion-mediated Proteolysis combined with an Electrophoretic Separation, permits to perform subsequently mAb tryptic digestion and electrophoresis separation of proteolysis products in an automated manner. Tryptic digestion conditions were optimized regarding the influence of enzyme concentration and incubation time in order to achieve similar enzymatic digestion efficiency to that obtained with the classical methodology (off-line). Then, the optimization of electrophoretic separation conditions concerning the nature of background electrolyte (BGE), ionic strength and pH was realized. Successful and repeatable electrophoretic profiles of three mAbs digests (Trastuzumab, Infliximab and Tocilizumab), comparable to the off-line digestion profiles, were obtained demonstrating the feasibility and robustness of the proposed methodology. In summary, the use of the proposed and optimized in-line approach opens a new, fast and easy way for the quality control of mAbs.
Url:
DOI: 10.1016/j.chroma.2017.10.066
PubMed: 29122286
Affiliations:
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pubmed:29122286Le document en format XML
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type="city">Montpellier</settlement></placeName></affiliation></author><author><name sortKey="Montels, Jerome" sort="Montels, Jerome" uniqKey="Montels J" first="Jérôme" last="Montels">Jérôme Montels</name><affiliation wicri:level="3"><nlm:affiliation>Institut des Biomolécules Max Mousseron-IBMM, Centre National de la Recherche Scientifique, Université de Montpellier, Unité Mixte de Recherche 5247, Faculté de Pharmacie, F-34093, Montpellier cedex 5, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Institut des Biomolécules Max Mousseron-IBMM, Centre National de la Recherche Scientifique, Université de Montpellier, Unité Mixte de Recherche 5247, Faculté de Pharmacie, F-34093, Montpellier cedex 5</wicri:regionArea><placeName><region type="region" nuts="2">Occitanie (région administrative)</region><region type="old region" nuts="2">Languedoc-Roussillon</region><settlement type="city">Montpellier</settlement></placeName></affiliation></author><author><name sortKey="Morel, Jacques" sort="Morel, Jacques" uniqKey="Morel J" first="Jacques" last="Morel">Jacques Morel</name><affiliation wicri:level="3"><nlm:affiliation>Département de Rhumatologie, Université de Montpellier, Hôpital Lapeyronie, 34295, Montpellier cedex 5, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Département de Rhumatologie, Université de Montpellier, Hôpital Lapeyronie, 34295, Montpellier cedex 5</wicri:regionArea><placeName><region type="region" nuts="2">Occitanie (région administrative)</region><region type="old region" nuts="2">Languedoc-Roussillon</region><settlement type="city">Montpellier</settlement></placeName></affiliation></author><author><name sortKey="Perrin, Catherine" sort="Perrin, Catherine" uniqKey="Perrin C" first="Catherine" last="Perrin">Catherine Perrin</name><affiliation wicri:level="3"><nlm:affiliation>Institut des Biomolécules Max Mousseron-IBMM, Centre National de la Recherche Scientifique, Université de Montpellier, Unité Mixte de Recherche 5247, Faculté de Pharmacie, F-34093, Montpellier cedex 5, France. Electronic address: catherine.perrin@umontpellier.fr.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Institut des Biomolécules Max Mousseron-IBMM, Centre National de la Recherche Scientifique, Université de Montpellier, Unité Mixte de Recherche 5247, Faculté de Pharmacie, F-34093, Montpellier cedex 5</wicri:regionArea><placeName><region type="region" nuts="2">Occitanie (région administrative)</region><region type="old region" nuts="2">Languedoc-Roussillon</region><settlement type="city">Montpellier</settlement></placeName></affiliation></author></analytic><series><title level="j">Journal of chromatography. A</title><idno type="eISSN">1873-3778</idno><imprint><date when="2017" type="published">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Monoclonal</term><term>Antibodies, Monoclonal, Humanized (analysis)</term><term>Automation</term><term>Chemistry, Pharmaceutical (instrumentation)</term><term>Electrophoresis, Capillary</term><term>Enzyme Assays (methods)</term><term>Hydrogen-Ion Concentration</term><term>Osmolar Concentration</term><term>Proteolysis</term><term>Quality Control</term><term>Trastuzumab (analysis)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Anticorps monoclonaux</term><term>Anticorps monoclonaux humanisés (analyse)</term><term>Automatisation</term><term>Chimie pharmaceutique (instrumentation)</term><term>Concentration en ions d'hydrogène</term><term>Concentration osmolaire</term><term>Contrôle de qualité</term><term>Dosages enzymatiques ()</term><term>Protéolyse</term><term>Trastuzumab (analyse)</term><term>Électrophorèse capillaire</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Antibodies, Monoclonal, Humanized</term><term>Trastuzumab</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Antibodies, Monoclonal</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Anticorps monoclonaux humanisés</term><term>Trastuzumab</term></keywords><keywords scheme="MESH" qualifier="instrumentation" xml:lang="en"><term>Chemistry, Pharmaceutical</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Enzyme Assays</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Automation</term><term>Electrophoresis, Capillary</term><term>Hydrogen-Ion Concentration</term><term>Osmolar Concentration</term><term>Proteolysis</term><term>Quality Control</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Anticorps monoclonaux</term><term>Automatisation</term><term>Chimie pharmaceutique</term><term>Concentration en ions d'hydrogène</term><term>Concentration osmolaire</term><term>Contrôle de qualité</term><term>Dosages enzymatiques</term><term>Protéolyse</term><term>Électrophorèse capillaire</term></keywords><keywords scheme="mix" xml:lang="en"><term> Antibodies</term><term> Automation</term><term> Capillary</term><term> Capillary electrophoresis</term><term> Chemistry</term><term> Electrophoresis</term><term> Enzymatic microreactor</term><term> Enzyme Assays</term><term> Humanized</term><term> Hydrogen-Ion Concentration</term><term> In-line digestion</term><term> Monoclonal</term><term> Monoclonal antibody</term><term> Osmolar Concentration</term><term> Pharmaceutical</term><term> Proteolysis</term><term> Proteolysis product separation</term><term> Quality control</term><term> Trastuzumab</term><term>Quality Control</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The main purpose of the present work is to provide a fully integrated miniaturized electrophoretic methodology in order to facilitate the quality control of monoclonal antibodies (mAbs). This methodology called D-PES, which stands for Diffusion-mediated Proteolysis combined with an Electrophoretic Separation, permits to perform subsequently mAb tryptic digestion and electrophoresis separation of proteolysis products in an automated manner. Tryptic digestion conditions were optimized regarding the influence of enzyme concentration and incubation time in order to achieve similar enzymatic digestion efficiency to that obtained with the classical methodology (off-line). Then, the optimization of electrophoretic separation conditions concerning the nature of background electrolyte (BGE), ionic strength and pH was realized. Successful and repeatable electrophoretic profiles of three mAbs digests (Trastuzumab, Infliximab and Tocilizumab), comparable to the off-line digestion profiles, were obtained demonstrating the feasibility and robustness of the proposed methodology. In summary, the use of the proposed and optimized in-line approach opens a new, fast and easy way for the quality control of mAbs.</div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Languedoc-Roussillon</li><li>Occitanie (région administrative)</li></region><settlement><li>Montpellier</li></settlement></list><tree><country name="France"><region name="Occitanie (région administrative)"><name sortKey="Ladner, Yoann" sort="Ladner, Yoann" uniqKey="Ladner Y" first="Yoann" last="Ladner">Yoann Ladner</name></region><name sortKey="Bartley, Killian" sort="Bartley, Killian" uniqKey="Bartley K" first="Killian" last="Bartley">Killian Bartley</name><name sortKey="Coussot, Gaelle" sort="Coussot, Gaelle" uniqKey="Coussot G" first="Gaelle" last="Coussot">Gaelle Coussot</name><name sortKey="Mas, Silvia" sort="Mas, Silvia" uniqKey="Mas S" first="Silvia" last="Mas">Silvia Mas</name><name sortKey="Montels, Jerome" sort="Montels, Jerome" uniqKey="Montels J" first="Jérôme" last="Montels">Jérôme Montels</name><name sortKey="Morel, Jacques" sort="Morel, Jacques" uniqKey="Morel J" first="Jacques" last="Morel">Jacques Morel</name><name sortKey="Perrin, Catherine" sort="Perrin, Catherine" uniqKey="Perrin C" first="Catherine" last="Perrin">Catherine Perrin</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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