| In vitro affinity maturation of human GM-CSF antibodies by targeted CDR-diversification. | |
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MedLine Citation:
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PMID: 18722015 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The mammalian immune system applies somatic hypermutation to select for antibodies with improved dissociation rates in vivo up to an intrinsic limit, previously termed as affinity ceiling. However, for certain therapeutic applications it may be desirable to further improve antibody affinities beyond that limit. In this study the selection of antibodies specific for the pro-inflammatory cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) from the HuCAL GOLD human antibody library is described. In order to increase affinity and also functional activity, in vitro affinity maturation of a pool of lead Fab candidates was carried out. CDR-L3 and parallel CDR-H2 diversification using trinucleotide consensus cassettes were followed by the combination of optimized CDR-L3 and CDR-H2 leading to a 5000-fold improved affinity finally reaching a K(D) of 400 fM. Cytokine neutralizing potential of MOR04357 was evaluated in a TF-1 proliferation assay. Along with affinity optimization a 2000-fold increase in potency was observed compared to the parental antibody. Due to species cross-reactivity MOR04357 also blocks rat GM-CSF induced proliferation of FDCP-1 cells. Receptor inhibition studies showed that MOR04357 prevents the interaction of GM-CSF with the GM-CSF receptor alpha chain. As a consequence this leads to a blockade in signal transduction as measured by abolished STAT5 phosphorylation in the presence of GM-CSF and antibody. Due to its pro-inflammatory role GM-CSF has been implicated in the pathophysiology of inflammatory diseases like rheumatoid arthritis or asthma. Based on the mode of action described herein MOR04357 shows favourable antibody features as a potential drug candidate. |
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Authors:
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Stefan Steidl; Olaf Ratsch; Bodo Brocks; Manuela Dürr; Elisabeth Thomassen-Wolf |
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Publication Detail:
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Type: Journal Article Date: 2008-08-21 |
Journal Detail:
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Title: Molecular immunology Volume: 46 ISSN: 0161-5890 ISO Abbreviation: Mol. Immunol. Publication Date: 2008 Nov |
Date Detail:
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Created Date: 2008-10-17 Completed Date: 2009-04-08 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 7905289 Medline TA: Mol Immunol Country: England |
Other Details:
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Languages: eng Pagination: 135-44 Citation Subset: IM |
Affiliation:
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MorphoSys AG, Lena-Christ-Str. 48, Martinsried 82152, Germany. steidl@morphosys.com |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Antibody Affinity / immunology* CHO Cells Cell Nucleus / metabolism Cell Proliferation Clone Cells Complementarity Determining Regions / immunology* Cricetinae Cricetulus Cross Reactions / immunology Granulocyte-Macrophage Colony-Stimulating Factor / immunology* Humans Immunoglobulin G / immunology Immunoglobulin Heavy Chains / immunology Immunoglobulin Light Chains / immunology Macaca mulatta Neutralization Tests Peptide Library Phosphorylation Protein Transport Rats Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / immunology STAT5 Transcription Factor / metabolism |
| Chemical | |
Reg. No./Substance:
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0/Complementarity Determining Regions; 0/Immunoglobulin G; 0/Immunoglobulin Heavy Chains; 0/Immunoglobulin Light Chains; 0/Peptide Library; 0/Receptors, Granulocyte-Macrophage Colony-Stimulating Factor; 0/STAT5 Transcription Factor; 83869-56-1/Granulocyte-Macrophage Colony-Stimulating Factor |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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