Rational design of a conformation-specific antibody for the quantification of Aβ oligomers
| Autor: | Ryan Limbocker, Tom Scheidt, Francesco A. Aprile, Michele Vendruscolo, Patricia C. Salinas, Pietro Sormanni, Michele Perni, Tuomas P. J. Knowles, Johnny Habchi, Christopher M. Dobson, Tomas Sneideris, Shianne Chhangur, Gabriella T. Heller, Francesco Simone Ruggeri, Steven F. Lee, Marina Podpolny, Lisa-Maria Needham, Benedetta Mannini |
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| Přispěvatelé: | Alzheimer's Society, Medical Research Council (MRC), Aprile, Francesco A [0000-0002-5040-4420], Sormanni, Pietro [0000-0002-6228-2221], Podpolny, Marina [0000-0003-2226-1183], Ruggeri, Francesco S [0000-0002-1232-1907], Perni, Michele [0000-0001-7593-8376], Scheidt, Tom [0000-0002-0185-7730], Mannini, Benedetta [0000-0001-6812-7348], Habchi, Johnny [0000-0003-4898-9623], Lee, Steven F [0000-0003-4492-5139], Vendruscolo, Michele [0000-0002-3616-1610], Apollo - University of Cambridge Repository |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Amyloid Protein Conformation Protein design Peptide Computational biology Protein aggregation Hippocampus Epitope Antibodies protein aggregation 03 medical and health sciences Epitopes Mice Protein Aggregates 0302 clinical medicine Antigen Alzheimer Disease Antibody Specificity Animals Caenorhabditis elegans protein design chemistry.chemical_classification Science & Technology Multidisciplinary Amyloid beta-Peptides Chemistry Rational design amyloid Alzheimer's disease Single-Domain Antibodies Biological Sciences Multidisciplinary Sciences ALZHEIMERS-DISEASE Disease Models Animal Biophysics and Computational Biology 030104 developmental biology Science & Technology - Other Topics Protein folding Target protein Alzheimer’s disease 030217 neurology & neurosurgery Protein Binding |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America Proceedings of the National Academy of Sciences of the United States of America, 117(24), 13509-13518 Proceedings of the National Academy of Sciences of the United States of America 117 (2020) 24 |
| ISSN: | 1091-6490 0027-8424 |
| Popis: | Significance The accurate quantification of the amounts of small oligomeric assemblies formed by the amyloid β (Aβ) peptide represents a major challenge in the Alzheimer’s field. There is therefore great interest in the development of methods to specifically detect these oligomers by distinguishing them from larger aggregates. The availability of these methods will enable the development of effective diagnostic and therapeutic interventions for this and other diseases related to protein misfolding and aggregation. We describe here a single-domain antibody able to selectively quantify oligomers of the Aβ peptide in isolation and in complex protein mixtures from animal models of disease. Protein misfolding and aggregation is the hallmark of numerous human disorders, including Alzheimer’s disease. This process involves the formation of transient and heterogeneous soluble oligomers, some of which are highly cytotoxic. A major challenge for the development of effective diagnostic and therapeutic tools is thus the detection and quantification of these elusive oligomers. Here, to address this problem, we develop a two-step rational design method for the discovery of oligomer-specific antibodies. The first step consists of an “antigen scanning” phase in which an initial panel of antibodies is designed to bind different epitopes covering the entire sequence of a target protein. This procedure enables the determination through in vitro assays of the regions exposed in the oligomers but not in the fibrillar deposits. The second step involves an “epitope mining” phase, in which a second panel of antibodies is designed to specifically target the regions identified during the scanning step. We illustrate this method in the case of the amyloid β (Aβ) peptide, whose oligomers are associated with Alzheimer’s disease. Our results show that this approach enables the accurate detection and quantification of Aβ oligomers in vitro, and in Caenorhabditis elegans and mouse hippocampal tissues. |
| Databáze: | OpenAIRE |
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