6b9y: Difference between revisions
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==Trastuzumab Fab v3 in complex with 5-phenyl meditope variant== | |||
<StructureSection load='6b9y' size='340' side='right' caption='[[6b9y]], [[Resolution|resolution]] 2.14Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[6b9y]] is a 5 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6B9Y OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6B9Y FirstGlance]. <br> | |||
</td></tr><tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=ACE:ACETYL+GROUP'>ACE</scene></td></tr> | |||
<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4ioi|4ioi]], [[5u3d|5u3d]]</td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6b9y FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6b9y OCA], [http://pdbe.org/6b9y PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6b9y RCSB], [http://www.ebi.ac.uk/pdbsum/6b9y PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6b9y ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
The high specificity and favorable pharmacological properties of monoclonal antibodies (mAbs) have prompted significant interest in re-engineering this class of molecules to add novel functionalities for enhanced therapeutic and diagnostic potential. Here, we used the high affinity, meditope-Fab interaction to template and drive the rapid, efficient, and stable site-specific formation of a disulfide bond. We demonstrate that this template-catalyzed strategy provides a consistent and reproducible means to conjugate fluorescent dyes, cytotoxins, or "click" chemistry handles to meditope-enabled mAbs (memAbs) and memFabs. More importantly, we demonstrate this covalent functionalization is achievable using natural amino acids only, opening up the opportunity to genetically encode cysteine meditope "tags" to biologics. As proof of principle, genetically encoded, cysteine meditope tags were added to the N- and/or C-termini of fluorescent proteins, nanobodies, and affibodies, each expressed in bacteria, purified to homogeneity, and efficiently conjugated to different memAbs and meFabs. We further show that multiple T-cell and Her2-targeting bispecific molecules using this strategy potently activate T-cell signaling pathways in vitro. Finally, the resulting products are highly stable as evidenced by serum stability assays (>14 d at 37 degrees C) and in vivo imaging of tumor xenographs. Collectively, the platform offers the opportunity to build and exchange an array of functional moieties, including protein biologics, among any cysteine memAb or Fab to rapidly create, test, and optimize stable, multifunctional biologics. | |||
Template-Catalyzed, Disulfide Conjugation of Monoclonal Antibodies Using a Natural Amino Acid Tag.,King JD, Ma Y, Kuo YC, Bzymek KP, Goodstein LH, Meyer K, Moore RE, Crow D, Colcher DM, Singh G, Horne DA, Williams JC Bioconjug Chem. 2018 Jun 20;29(6):2074-2081. doi:, 10.1021/acs.bioconjchem.8b00284. Epub 2018 May 25. PMID:29763554<ref>PMID:29763554</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
[[Category: | <div class="pdbe-citations 6b9y" style="background-color:#fffaf0;"></div> | ||
[[Category: Williams, J | == References == | ||
[[Category: | <references/> | ||
__TOC__ | |||
</StructureSection> | |||
[[Category: Bzymek, K P]] | |||
[[Category: King, J D]] | |||
[[Category: Williams, J C]] | |||
[[Category: Fab]] | |||
[[Category: Immune system]] | |||
[[Category: Meditope]] | |||
[[Category: Monoclonal antibody]] | |||
Revision as of 13:04, 5 September 2018
Trastuzumab Fab v3 in complex with 5-phenyl meditope variantTrastuzumab Fab v3 in complex with 5-phenyl meditope variant
Structural highlights
Publication Abstract from PubMedThe high specificity and favorable pharmacological properties of monoclonal antibodies (mAbs) have prompted significant interest in re-engineering this class of molecules to add novel functionalities for enhanced therapeutic and diagnostic potential. Here, we used the high affinity, meditope-Fab interaction to template and drive the rapid, efficient, and stable site-specific formation of a disulfide bond. We demonstrate that this template-catalyzed strategy provides a consistent and reproducible means to conjugate fluorescent dyes, cytotoxins, or "click" chemistry handles to meditope-enabled mAbs (memAbs) and memFabs. More importantly, we demonstrate this covalent functionalization is achievable using natural amino acids only, opening up the opportunity to genetically encode cysteine meditope "tags" to biologics. As proof of principle, genetically encoded, cysteine meditope tags were added to the N- and/or C-termini of fluorescent proteins, nanobodies, and affibodies, each expressed in bacteria, purified to homogeneity, and efficiently conjugated to different memAbs and meFabs. We further show that multiple T-cell and Her2-targeting bispecific molecules using this strategy potently activate T-cell signaling pathways in vitro. Finally, the resulting products are highly stable as evidenced by serum stability assays (>14 d at 37 degrees C) and in vivo imaging of tumor xenographs. Collectively, the platform offers the opportunity to build and exchange an array of functional moieties, including protein biologics, among any cysteine memAb or Fab to rapidly create, test, and optimize stable, multifunctional biologics. Template-Catalyzed, Disulfide Conjugation of Monoclonal Antibodies Using a Natural Amino Acid Tag.,King JD, Ma Y, Kuo YC, Bzymek KP, Goodstein LH, Meyer K, Moore RE, Crow D, Colcher DM, Singh G, Horne DA, Williams JC Bioconjug Chem. 2018 Jun 20;29(6):2074-2081. doi:, 10.1021/acs.bioconjchem.8b00284. Epub 2018 May 25. PMID:29763554[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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