Abstract: TH-PO0357
New Antibody Tools to Study Alport Syndrome
Session Information
- Glomerular Diseases: Genetics to Therapeutics
October 22, 2026 | Location: Exhibit Hall A, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology
Authors
- Fresquet, Maryline, The University of Manchester Faculty of Biology Medicine and Health, Manchester, England, United Kingdom
- Bolas, Gema, The University of Manchester Faculty of Biology Medicine and Health, Manchester, England, United Kingdom
- Kaseda, Shota, The University of Manchester Faculty of Biology Medicine and Health, Manchester, England, United Kingdom
- Williams, Emily, The University of Manchester Faculty of Biology Medicine and Health, Manchester, England, United Kingdom
- Newton, Sherylanne, University College London, London, England, United Kingdom
- Sugden, Conor J., The University of Manchester Faculty of Biology Medicine and Health, Manchester, England, United Kingdom
- Jowitt, Thomas A., The University of Manchester Faculty of Biology Medicine and Health, Manchester, England, United Kingdom
- Goncalves, Kevin A., Deerfield Management Company LP, New York, New York, United States
- Steinsapir, Andrew, Deerfield Management Company LP, New York, New York, United States
- Jagger, Dan, University College London, London, England, United Kingdom
- Sherwood, David R., Duke University, Durham, North Carolina, United States
- Lennon, Rachel, The University of Manchester Faculty of Biology Medicine and Health, Manchester, England, United Kingdom
Background
Type IV collagen is essential for normal basement membrane structure and function and exists as three trimeric alpha chain networks: α1α1α2, α3α4α5 and α5α5α6. Variants in COL4A3, COL4A4, and COL4A5 cause Alport syndrome and disrupt the formation of the α3α4α5 network, leading to abnormalities in the kidney, cochlea, and eye. Here, we developed and characterised new mouse monoclonal antibodies targeting a human mini-α3α4α5 heterotrimer.
Methods
We generated three monoclonal antibodies and tested their ability to bind monomeric alpha-3, alpha-5, and the α3α4α5 trimer. We assessed binding specificity and affinity using slot blotting, western blotting, and surface plasmon resonance. We tested the antibodies in human and mouse tissues using immunofluorescence and examined their specificity in knockout mouse models lacking Col4a3, Col4a5, or Col4a6. We also performed immunoprecipitation with the antibodies to isolate COL4A345 from solubilised glomeruli.
Results
We found that the antigen used to generate the antibodies formed a stable mini-α3α4α5 trimer. Clone #72 bound an epitope involving both alpha-3 and alpha-5, while clones #5 and #44 recognised only the intact α3α4α5 complex. All antibodies were highly specific for α3α4α5 and did not cross-react with the other collagen IV networks α1α1α2 or α5α5α6. In tissue immunofluorescence, they showed strong localisation to basement membranes in kidney, cochlea, and eye tissues from humans and mice.
Conclusion
Clone #72 performed best overall, showing strong and specific detection collagen IV-α3α4α5 in tissues. These antibodies provide useful new tools for studying collagen IV and will help improve detection of intact α3α4α5. in clinical and research settings.
Funding
- Commercial Support – Deerfield Management Company