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Isolating Anti-amyloid Antibodies from Yeast-Displayed Libraries
- Source :
- Methods in Molecular Biology ISBN: 9781071622841, Methods Mol Biol
- Publication Year :
- 2022
- Publisher :
- Springer US, 2022.
-
Abstract
- Conformational antibodies specific for amyloid-forming peptides and proteins are important for a range of biomedical applications, including detecting, inhibiting, and potentially treating protein aggregation disorders ranging from Alzheimer's to Parkinson's diseases. Generation of anti-amyloid antibodies is greatly complicated by the complex, heterogeneous and insoluble nature of amyloid antigens. Here we describe systematic methods for isolating and affinity maturing anti-amyloid antibodies using yeast surface display. Magnetic-activated cell sorting is used to sort single-chain antibody libraries positively for binding to amyloid antigens and negatively against the corresponding disaggregated antigens to remove antibodies that bind in a conformation-independent manner. Isolated lead antibody clones with conformational specificity are affinity matured via targeted CDR mutagenesis and magnetic-activated cell sorting.
Details
- ISBN :
- 978-1-07-162284-1
- ISBNs :
- 9781071622841
- Database :
- OpenAIRE
- Journal :
- Methods in Molecular Biology ISBN: 9781071622841, Methods Mol Biol
- Accession number :
- edsair.doi.dedup.....63dee8dc620e4a18f26c56bb7f2d799a