Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries

View/ Open
Issue Date
2022-06-05Author
Huang, Aric
Jin, Wei
Fahad, Ahmed S.
Mussman, Brooklyn K.
Nicchia, Grazia Paola
Madan, Bharat
de Souza, Matheus Oliveira
Griffin, J. Daniel
Bennett, Jeffrey L.
Frigeri, Antonio
Berkland, Cory J.
DeKosky, Brandon J.
Publisher
MDPI
Type
Article
Article Version
Scholarly/refereed, publisher version
Rights
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license.
Metadata
Show full item recordAbstract
A rapid and effective method to identify disease-specific antibodies from clinical patients is important for understanding autoimmune diseases and for the development of effective disease therapies. In neuromyelitis optica (NMO), the identification of antibodies targeting the aquaporin-4 (AQP4) membrane protein traditionally involves the labor-intensive and time-consuming process of single B-cell sorting, followed by antibody cloning, expression, purification, and analysis for anti-AQP4 activity. To accelerate patient-specific antibody discovery, we compared two unique approaches for screening anti-AQP4 antibodies from yeast antibody surface display libraries. Our first approach, cell-based biopanning, has strong advantages for its cell-based display of native membrane-bound AQP4 antigens and is inexpensive and simple to perform. Our second approach, FACS screening using solubilized AQP4 antigens, permits real-time population analysis and precision sorting for specific antibody binding parameters. We found that both cell-based biopanning and FACS screening were effective for the enrichment of AQP4-binding clones. These screening techniques will enable library-scale functional interrogation of large natively paired antibody libraries for comprehensive analysis of anti-AQP4 antibodies in clinical samples and for robust therapeutic discovery campaigns.
Collections
Citation
Huang, A.; Jin, W.; Fahad, A.S.; Mussman, B.K.; Nicchia, G.P.; Madan, B.; de Souza, M.O.; Griffin, J.D.; Bennett, J.L.; Frigeri, A.; Berkland, C.J.; DeKosky, B.J. Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries. Antibodies 2022, 11, 39. https://doi.org/10.3390/antib11020039
Items in KU ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.
We want to hear from you! Please share your stories about how Open Access to this item benefits YOU.