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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Sormanni, Pietro Aprile, Francesco A. Vendruscolo, Michele |
| Description | Author Affiliation: Sormanni P ( Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom.); Aprile FA ( Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom.); Vendruscolo M ( Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom mv245@cam.ac.uk.); |
| Abstract | Antibodies are powerful tools in life sciences research, as well as in diagnostic and therapeutic applications, because of their ability to bind given molecules with high affinity and specificity. Using current methods, however, it is laborious and sometimes difficult to generate antibodies to target specific epitopes within a protein, in particular if these epitopes are not effective antigens. Here we present a method to rationally design antibodies to enable them to bind virtually any chosen disordered epitope in a protein. The procedure consists in the sequence-based design of one or more complementary peptides targeting a selected disordered epitope and the subsequent grafting of such peptides on an antibody scaffold. We illustrate the method by designing six single-domain antibodies to bind different epitopes within three disease-related intrinsically disordered proteins and peptides ( -synuclein, Aß42, and IAPP). Our results show that all these designed antibodies bind their targets with good affinity and specificity. As an example of an application, we show that one of these antibodies inhibits the aggregation of -synuclein at substoichiometric concentrations and that binding occurs at the selected epitope. Taken together, these results indicate that the design strategy that we propose makes it possible to obtain antibodies targeting given epitopes in disordered proteins or protein regions. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 32 |
| Volume Number | 112 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2015-08-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Antibodies Chemistry Epitopes Immunology Intrinsically Disordered Proteins Protein Engineering Amino Acid Sequence Amyloid Beta-Peptides Antibody Specificity Blotting, Western Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Fluorescence Islet Amyloid Polypeptide Molecular Sequence Data Mutant Proteins Peptides Protein Aggregates Protein Binding Protein Structure, Secondary Single-Domain Antibodies Alpha-Synuclein Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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