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Lock-and-key motif as a concept for designing affinity adsorbents for protein purification

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dc.contributor.author Platis, D en
dc.contributor.author Sotriffer, CA en
dc.contributor.author Clonis, Y en
dc.contributor.author Labrou, NE en
dc.date.accessioned 2014-06-06T06:47:09Z
dc.date.available 2014-06-06T06:47:09Z
dc.date.issued 2006 en
dc.identifier.issn 00219673 en
dc.identifier.uri http://dx.doi.org/10.1016/j.chroma.2006.06.051 en
dc.identifier.uri http://62.217.125.90/xmlui/handle/123456789/3420
dc.subject Affinity chromatography en
dc.subject Antibody purification en
dc.subject Combinatorial design en
dc.subject Ligand design en
dc.subject Molecular modeling en
dc.subject.other Antibody purification en
dc.subject.other Combinatorial design en
dc.subject.other Ligand design en
dc.subject.other Molecular modeling en
dc.subject.other Adsorbents en
dc.subject.other Affinity chromatography en
dc.subject.other Antibodies en
dc.subject.other Immunology en
dc.subject.other Molecular dynamics en
dc.subject.other Purification en
dc.subject.other Synthesis (chemical) en
dc.subject.other Proteins en
dc.subject.other glutathione transferase en
dc.subject.other adsorption en
dc.subject.other article en
dc.subject.other binding affinity en
dc.subject.other bioinformatics en
dc.subject.other ligand binding en
dc.subject.other molecular dynamics en
dc.subject.other molecular model en
dc.subject.other molecular recognition en
dc.subject.other priority journal en
dc.subject.other protein purification en
dc.subject.other purification en
dc.subject.other solid en
dc.subject.other synthesis en
dc.subject.other Amino Acid Motifs en
dc.subject.other Amino Acid Sequence en
dc.subject.other Binding Sites en
dc.subject.other Biomimetics en
dc.subject.other Chromatography, Affinity en
dc.subject.other Combinatorial Chemistry Techniques en
dc.subject.other Dipeptides en
dc.subject.other Glutathione Transferase en
dc.subject.other HIV en
dc.subject.other HIV Antibodies en
dc.subject.other Humans en
dc.subject.other Ligands en
dc.subject.other Models, Molecular en
dc.subject.other Molecular Sequence Data en
dc.subject.other Protein Binding en
dc.subject.other Protein Subunits en
dc.subject.other Recombinant Proteins en
dc.subject.other Seeds en
dc.subject.other Structure-Activity Relationship en
dc.subject.other Triazines en
dc.subject.other Zea mays en
dc.subject.other Human immunodeficiency virus 2 en
dc.subject.other Zea mays en
dc.title Lock-and-key motif as a concept for designing affinity adsorbents for protein purification en
heal.type journalArticle en
heal.identifier.primary 10.1016/j.chroma.2006.06.051 en
heal.publicationDate 2006 en
heal.abstract The lock-and-key (LAK) motif, a common structural moiety found in subunit interfaces of glutathione S-transferases (GSTs), plays an important role in biomolecular recognition and quaternary structure integrity. Inspection of the key structural features of the LAK motif prompted the de novo design and combinatorial synthesis of a 13-membered solid-phase ligand library, employing as a lead ligand the Phe-Trz-X structure, mimicking the LAK motif. 1,3,5-Triazine (Trz) was used as the scaffold for assembly, substituted with different LAK-mimetic amino acids. De novo ligand design was effected using bioinformatics and molecular modeling and based on mimicking the interactions of the LAK motif. The library of affinity adsorbents was assessed for binding corn and human serum proteomes and purified proteins of different structure and ligand binding specificity. The results showed remarkable differences in the binding specificity of LAK-mimetic adsorbents for a wide range of proteins, as a consequence of minor changes in ligand structure. One LAK-mimetic adsorbent was integrated in a single-step purification protocol for human monoclonal anti-human immunodeficiency virus 2F5 antibody (mAb 2F5) from spiked corn extract, affording high recovery and purity. The results demonstrate that the principle of natural recognition found in the lock-and-key motif, in combination with de novo combinatorial design, may lead to synthetic affinity ligands, useful in downstream processing and proteomic research. © 2006 Elsevier B.V. All rights reserved. en
heal.journalName Journal of Chromatography A en
dc.identifier.issue 1-2 en
dc.identifier.volume 1128 en
dc.identifier.doi 10.1016/j.chroma.2006.06.051 en
dc.identifier.spage 138 en
dc.identifier.epage 151 en


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