Strategies to Screen Anti-AQP4 Antibodies from Yeast Surface Display Libraries
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
Affiliations
Aric Huang
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USA
Wei Jin
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USA
Ahmed S. Fahad
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USA
Brooklyn K. Mussman
Department of Chemical Engineering, The University of Kansas, Lawrence, KS 66044, USA
Grazia Paola Nicchia
Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari Aldo Moro, 70121 Bari, Italy
Bharat Madan
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USA
Matheus Oliveira de Souza
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USA
J. Daniel Griffin
Bioengineering Graduate Program, The University of Kansas, Lawrence, KS 66045, USA
Jeffrey L. Bennett
Departments of Neurology and Ophthalmology, Programs in Neuroscience and Immunology, University of Colorado at Anschutz Medical Campus, Aurora, CO 80045, USA
Antonio Frigeri
Department of Basic Medical Sciences, Neurosciences and Sense Organs, School of Medicine, University of Bari Aldo Moro, 70121 Bari, Italy
Cory J. Berkland
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USA
Brandon J. DeKosky
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS 66044, USA
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.
