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Abstract: SA-PO349

Identification of Novel Target Antigens in Sera of Membranous Nephropathy Patients Using Whole Proteome Peptide Array Technology

Session Information

Category: Glomerular Diseases

  • 1202 Glomerular Diseases: Immunology and Inflammation

Authors

  • Prunotto, Marco, F. Hoffmann-La Roche Ltd., Basel, Switzerland
  • Bruschi, Maurizio, Laboratory of Molecular Nephrology, Istituto Giannina Gaslini, Genoa, Italy
  • Cavalli, Andrea, Universita della Svizzera Italiana, Bellinzona, Switzerland
  • Tan, John C., Roche Diagnostics, Madison, Wisconsin, United States
  • Candiano, Giovanni, Laboratory of Molecular Nephrology, Istituto Giannina Gaslini, Genova, Italy, Genoa, Italy
  • Mcclintock, Dana, Genentech, Inc, South San Francisco, California, United States
  • Patel, Jigar, Pharmaceutical, Madison, Wisconsin, United States
  • Ghiggeri, Gian Marco, Istituto Giannina Gaslini, Genoa, Italy
Background

Membranous nephropathy (MN) autoantigens have been discovered using the established laboratory technique of Western blotting, in which human sera from patients with MN were utilized to screen for specific reactivity with proteins extracted from human glomerular fractions. The objective of the present study was to investigate a highly characterized group of patients using a technology routinely adopted in antibody epitope mapping.

Methods

Sera from patients with membranous nephropathy, selected to have good (n=5) or bad prognosis (n=5) at hospitalization (T0) or twelve months after (T12) were hybridized with a whole proteome peptide array chip. The intensity of the signal retrieved by whole proteome peptide array chip was used to feed a weight gene co-expression network analysis (WGCNA) algorithm to identify a network of relationships between the signals and renal outcome in search of established and novel antigens of MN.

Results

Whole proteome peptide array technology allowed retrieving known antigens (e.g., PLA2R1 or THSPD7A) and identified specific peptide sequences on those proteins recognized by autoantibodies predictive of renal outcome. Peptides belonging to previously unknown target antigens in MN were also retrieved.

Conclusion

Whole proteome peptide array and extracellular protein microarray technologies can be profitably applied to the study of autoimmune diseases. The technologies identify known antigens of MN and novel unknown ones. Those preliminary findings need extensive validation in a large patient set.