Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries
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) memb...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-06-01
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| Series: | Antibodies |
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| Online Access: | https://www.mdpi.com/2073-4468/11/2/39 |
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| author | Aric Huang Wei Jin Ahmed S. Fahad Brooklyn K. Mussman Grazia Paola Nicchia Bharat Madan Matheus Oliveira de Souza J. Daniel Griffin Jeffrey L. Bennett Antonio Frigeri Cory J. Berkland Brandon J. DeKosky |
| author_facet | Aric Huang Wei Jin Ahmed S. Fahad Brooklyn K. Mussman Grazia Paola Nicchia Bharat Madan Matheus Oliveira de Souza J. Daniel Griffin Jeffrey L. Bennett Antonio Frigeri Cory J. Berkland Brandon J. DeKosky |
| author_sort | Aric Huang |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-10T00:35:18Z |
| format | Article |
| id | doaj.art-59f3a6dc89844b95ad3e0160581a8e51 |
| institution | Directory Open Access Journal |
| issn | 2073-4468 |
| language | English |
| last_indexed | 2024-03-10T00:35:18Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibodies |
| spelling | doaj.art-59f3a6dc89844b95ad3e0160581a8e512023-11-23T15:18:22ZengMDPI AGAntibodies2073-44682022-06-011123910.3390/antib11020039Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display LibrariesAric Huang0Wei Jin1Ahmed S. Fahad2Brooklyn K. Mussman3Grazia Paola Nicchia4Bharat Madan5Matheus Oliveira de Souza6J. Daniel Griffin7Jeffrey L. Bennett8Antonio Frigeri9Cory J. Berkland10Brandon J. DeKosky11Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USADepartment of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USADepartment of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USADepartment of Chemical Engineering, The University of Kansas, Lawrence, KS 66044, USADepartment of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari Aldo Moro, 70121 Bari, ItalyDepartment of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USADepartment of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USABioengineering Graduate Program, The University of Kansas, Lawrence, KS 66045, USADepartments of Neurology and Ophthalmology, Programs in Neuroscience and Immunology, University of Colorado at Anschutz Medical Campus, Aurora, CO 80045, USADepartment of Basic Medical Sciences, Neurosciences and Sense Organs, School of Medicine, University of Bari Aldo Moro, 70121 Bari, ItalyDepartment of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USADepartment of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USAA 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.https://www.mdpi.com/2073-4468/11/2/39yeast displayantibody discoveryyeast library screeningbiopanningaquaporin-4 |
| spellingShingle | Aric Huang Wei Jin Ahmed S. Fahad Brooklyn K. Mussman Grazia Paola Nicchia Bharat Madan Matheus Oliveira de Souza J. Daniel Griffin Jeffrey L. Bennett Antonio Frigeri Cory J. Berkland Brandon J. DeKosky Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries Antibodies yeast display antibody discovery yeast library screening biopanning aquaporin-4 |
| title | Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries |
| title_full | Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries |
| title_fullStr | Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries |
| title_full_unstemmed | Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries |
| title_short | Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries |
| title_sort | strategies to screen anti aqp4 antibodies from yeast surface display libraries |
| topic | yeast display antibody discovery yeast library screening biopanning aquaporin-4 |
| url | https://www.mdpi.com/2073-4468/11/2/39 |
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