دورية أكاديمية
Designing Fcabs: well-expressed and stable high affinity antigen-binding Fc fragments.
| العنوان: | Designing Fcabs: well-expressed and stable high affinity antigen-binding Fc fragments. |
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| المؤلفون: | Wozniak-Knopp G; Christian Doppler Laboratory for Innovative Immunotherapeutics, Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU), Vienna, Muthgasse 18, 1190 Vienna, Austria., Stadlmayr G; Christian Doppler Laboratory for Innovative Immunotherapeutics, Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU), Vienna, Muthgasse 18, 1190 Vienna, Austria., Perthold JW; Institute of Molecular Modeling and Simulation, Department of Material Sciences, University of Natural Resources and Life Sciences (BOKU), Vienna, Muthgasse 18, 1190 Vienna, Austria., Stadlbauer K; Christian Doppler Laboratory for Innovative Immunotherapeutics, Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU), Vienna, Muthgasse 18, 1190 Vienna, Austria., Woisetschläger M, Sun H, Rüker F; Christian Doppler Laboratory for Innovative Immunotherapeutics, Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU), Vienna, Muthgasse 18, 1190 Vienna, Austria. |
| المصدر: | Protein engineering, design & selection : PEDS [Protein Eng Des Sel] 2017 Sep 01; Vol. 30 (9), pp. 657-671. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Oxford University Press Country of Publication: England NLM ID: 101186484 Publication Model: Print Cited Medium: Internet ISSN: 1741-0134 (Electronic) Linking ISSN: 17410126 NLM ISO Abbreviation: Protein Eng Des Sel Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Oxford, UK : Oxford University Press, c2003- |
| مواضيع طبية MeSH: | Antibodies, Monoclonal/*chemistry , Immunoglobulin Fc Fragments/*chemistry , Immunoglobulin G/*chemistry , Protein Engineering/*methods , Vascular Endothelial Growth Factor A/*antagonists & inhibitors, Amino Acid Sequence ; Antibodies, Monoclonal/biosynthesis ; Antibodies, Monoclonal/genetics ; Antibodies, Monoclonal/pharmacology ; Antigens/chemistry ; Antigens/genetics ; Antigens/immunology ; Bevacizumab/chemistry ; Bevacizumab/pharmacology ; Binding Sites ; Cell Proliferation/drug effects ; Cell Surface Display Techniques ; Gene Expression ; Human Umbilical Vein Endothelial Cells/cytology ; Human Umbilical Vein Endothelial Cells/drug effects ; Human Umbilical Vein Endothelial Cells/immunology ; Humans ; Immunoglobulin Fc Fragments/biosynthesis ; Immunoglobulin Fc Fragments/genetics ; Immunoglobulin Fc Fragments/pharmacology ; Immunoglobulin G/biosynthesis ; Immunoglobulin G/genetics ; Immunoglobulin G/pharmacology ; Models, Molecular ; Molecular Dynamics Simulation ; Protein Binding ; Protein Stability ; Protein Structure, Secondary ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/chemistry ; Recombinant Proteins/genetics ; Recombinant Proteins/pharmacology ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae/metabolism ; Vascular Endothelial Growth Factor A/chemistry ; Vascular Endothelial Growth Factor A/genetics ; Vascular Endothelial Growth Factor A/immunology |
| مستخلص: | Fc fragment with antigen-binding (Fcab) is a novel construct which can be selected to recognize specifically a wide variety of target proteins. We describe the selection and affinity maturation of Fcab clones targeting VEGF, an important pro-angiogenesis factor. To investigate the extent of engineering permissible to Fcabs we applied targeted mutagenesis to all three C-terminal loop structures and the C-terminus of the CH3 domain to isolate high-affinity binders by directed evolution and yeast display. The matured clone, CT6, binds to VEGF with low nanomolar affinity and inhibits VEGF-stimulated proliferation of human umbilical vein endothelial cells in vitro. Molecular dynamics simulations were performed to address flexibility of the molecular structure of CT6 and to approximate a structural ensemble in aqueous solution. Significantly higher RMSF levels of CT6 in comparison to wild-type Fc were limited to the elongated CD-loop in the CH3 domain, while the overall structural integrity was retained. This allowed the Fcab to replace the Fc portion of a mAb, in which both the CH3 and Fab are capable of antigen engagement: a construct called mAb2 was assembled with CT6 and the Fab of bevacizumab. This bispecific molecule showed more potent antagonistic activity than bevacizumab in vitro. Further evaluation for the potential of the CT6 Fcab in targeted therapy is warranted due to the possibility of being combined with other therapeutically meaningful targets. (© The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.) |
| فهرسة مساهمة: | Keywords: Fcab; VEGF; directed evolution; molecular dynamics; yeast display |
| المشرفين على المادة: | 0 (Antibodies, Monoclonal) 0 (Antigens) 0 (Immunoglobulin Fc Fragments) 0 (Immunoglobulin G) 0 (Recombinant Proteins) 0 (VEGFA protein, human) 0 (Vascular Endothelial Growth Factor A) 2S9ZZM9Q9V (Bevacizumab) |
| تواريخ الأحداث: | Date Created: 20171006 Date Completed: 20171122 Latest Revision: 20201209 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1093/protein/gzx042 |
| PMID: | 28981753 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1741-0134 |
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| DOI: | 10.1093/protein/gzx042 |