دورية أكاديمية
Comprehensive interrogation of a minimalist synthetic CDR-H3 library and its ability to generate antibodies with therapeutic potential.
| العنوان: | Comprehensive interrogation of a minimalist synthetic CDR-H3 library and its ability to generate antibodies with therapeutic potential. |
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| المؤلفون: | Mahon CM; Pfizer, Global Biotherapeutics Technologies, Grange Castle Business Park, Clondalkin, Dublin 22, Ireland., Lambert MA, Glanville J, Wade JM, Fennell BJ, Krebs MR, Armellino D, Yang S, Liu X, O'Sullivan CM, Autin B, Oficjalska K, Bloom L, Paulsen J, Gill D, Damelin M, Cunningham O, Finlay WJ |
| المصدر: | Journal of molecular biology [J Mol Biol] 2013 May 27; Vol. 425 (10), pp. 1712-30. Date of Electronic Publication: 2013 Feb 19. |
| نوع المنشور: | Journal Article; Randomized Controlled Trial |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 2985088R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1089-8638 (Electronic) Linking ISSN: 00222836 NLM ISO Abbreviation: J Mol Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Amsterdam : Elsevier Original Publication: 1959- : London : Academic Press |
| مواضيع طبية MeSH: | Antibody Specificity*/genetics , Peptide Library*, Complementarity Determining Regions/*immunology , Complementarity Determining Regions/*therapeutic use , Single-Chain Antibodies/*biosynthesis , Single-Chain Antibodies/*therapeutic use, Adaptor Proteins, Signal Transducing ; Animals ; Calcium-Binding Proteins ; Cloning, Molecular ; Complementarity Determining Regions/genetics ; HEK293 Cells ; Humans ; Intercellular Signaling Peptides and Proteins/genetics ; Intercellular Signaling Peptides and Proteins/immunology ; Mice ; Models, Molecular ; Mutation ; Receptor, Notch1/antagonists & inhibitors ; Receptor, Notch1/genetics ; Receptor, Notch1/immunology ; Single-Chain Antibodies/genetics |
| مستخلص: | We have generated large libraries of single-chain Fv antibody fragments (>10(10) transformants) containing unbiased amino acid diversity that is restricted to the central combining site of the stable, well-expressed DP47 and DPK22 germline V-genes. Library WySH2A was constructed to examine the potential for synthetic complementarity-determining region (CDR)-H3 diversity to act as the lone source of binding specificity. Library WySH2B was constructed to assess the necessity for diversification in both the H3 and L3. Both libraries provided diverse, specific antibodies, yielding a total of 243 unique hits against 7 different targets, but WySH2B produced fewer hits than WySH2A when selected in parallel. WySH2A also consistently produced hits of similar quality to WySH2B, demonstrating that the diversification of the CDR-L3 reduces library fitness. Despite the absence of deliberate bias in the library design, CDR length was strongly associated with the number of hits produced, leading to a functional loop length distribution profile that mimics the biases observed in the natural repertoire. A similar trend was also observed for the CDR-L3. After target selections, several key amino acids were enriched in the CDR-H3 (e.g., small and aromatic residues) while others were reduced (e.g., strongly charged residues) in a manner that was specific to position, preferentially occurred in CDR-H3 stem positions, and tended towards residues associated with loop stabilization. As proof of principle for the WySH2 libraries to produce viable lead candidate antibodies, 114 unique hits were produced against Delta-like ligand 4 (DLL4). Leads exhibited nanomolar binding affinities, highly specific staining of DLL4+ cells, and biochemical neutralization of DLL4-NOTCH1 interaction. (Copyright © 2013 Elsevier Ltd. All rights reserved.) |
| المشرفين على المادة: | 0 (Adaptor Proteins, Signal Transducing) 0 (Calcium-Binding Proteins) 0 (Complementarity Determining Regions) 0 (DLL4 protein, human) 0 (Intercellular Signaling Peptides and Proteins) 0 (NOTCH1 protein, human) 0 (Peptide Library) 0 (Receptor, Notch1) 0 (Single-Chain Antibodies) |
| تواريخ الأحداث: | Date Created: 20130223 Date Completed: 20130709 Latest Revision: 20201209 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1016/j.jmb.2013.02.015 |
| PMID: | 23429058 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1089-8638 |
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| DOI: | 10.1016/j.jmb.2013.02.015 |