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.
CITATION STYLE
Desai, A. A., Zupancic, J. M., Smith, M. D., & Tessier, P. M. (2022). Isolating Anti-amyloid Antibodies from Yeast-Displayed Libraries. In Methods in Molecular Biology (Vol. 2491, pp. 471–490). Humana Press Inc. https://doi.org/10.1007/978-1-0716-2285-8_22
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