Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries
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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jun 05, 2022
- Source ID
- 10.3390/antib11020039
Entities
People
- Ahmed S. Fahad
- Antonio Frigeri
- Aric Huang
- Bharat Madan
- Brandon J DeKosky
- Brooklyn K. Mussman
- Cory J. Berkland
- Grazia Paola Nicchia
- J. Daniel Griffin
- Jeffrey L. Bennett
- Matheus Oliveira De Souza
- Wei Jin
Organizations
- Air Force Office of Scientific Research
- National Institute of Allergy and Infectious Diseases
- National Institutes of Health