Isolating Anti-amyloid Antibodies from Yeast-Displayed Libraries

Autor:	Alec A, Desai, Jennifer M, Zupancic, Matthew D, Smith, Peter M, Tessier
Rok vydání:	2022
Předmět:	Amyloid Protein Aggregates Amyloidogenic Proteins Saccharomyces cerevisiae Antibodies Article
Zdroj:	Methods in Molecular Biology ISBN: 9781071622841 Methods Mol Biol
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::63dee8dc620e4a18f26c56bb7f2d799a https://doi.org/10.1007/978-1-0716-2285-8_22 Zobrazit plný text záznamu