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Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries
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 ... show 2 more
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
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Abstract
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.
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Date
2022-06-05
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Publisher
MDPI
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Keywords
Yeast display, Antibody discovery, Yeast library screening, Biopanning, Aquaporin-4
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