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E-Poster Submission Deadline
Please prepare and upload your E-Poster no later than March 14, 2026 11.59PM CET. After this date, you will no longer be able to prepare and upload your E-poster and it will not be displayed and accessible on the congress website.
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Abstract titles should be brief and reflect the content of the abstract.
Dysregulated complement activation contributes to glomerular injury in atypical hemolytic uremic syndrome, C3 glomerulopathy, diabetic kidney disease, and other glomerulonephritides. Conventional ELISAs quantify single factors or autoantibodies but are limited by low throughput, labor-intensive workflows, suboptimal specificity, and high sample consumption.
Figure 1. Schematic diagram of the ADDT-PEA detection principle
We developed an Antigen- and Antibody Dual-Target Proximity Extension Assay (AADT-PEA) for single-tube, multiplex measurement of 11 complement-related analytes. Seven soluble factors (sC5b-9, CFH, CFP, CFI, C3a, C5a, Ba) were each detected by two non-competing, factor-specific antibodies, with each antibody covalently conjugated to a distinct, barcode-encoded oligonucleotide; simultaneous binding brought the oligonucleotides into proximity, enabling hybridization and polymerase-mediated extension to generate target-specific amplicons quantified by next-generation sequencing (NGS). Four autoantibodies (anti-CFH, anti-CFI, anti-C3b, anti-factor B) were measured using the paired antigen–oligo and anti-IgG–oligo strategy, with proximity-triggered extension and NGS readout. Seven-point serial dilutions of positive standards were run for each analyte, and performance was compared with commercial ELISAs.
In patients with confirmed complement dysregulation, AADT-PEA demonstrated high analytical sensitivity and specificity. Calibration analyses showed excellent linearity across all 11 analytes (R²: 0.9984, 0.9976, 0.9968, 0.9991, 0.9993, 0.9989, 0.9994, 0.9979, 0.9985, 0.9990, 0.9989), exceeding commercial ELISAs (R²: 0.9503, 0.9742, 0.9655, 0.9568, 0.9724, 0.9603, 0.9458, 0.9871, 0.9394, 0.9506, 0.9477). Limits of detection were significantly lower than ELISA for all antibodies (P < 0.05), indicating superior sensitivity.
AADT-PEA enables high-throughput, multiplex quantification of complement factors and autoantibodies from minimal sample input in a single reaction, offering improved accuracy and sensitivity relative to ELISA and reduced cross-reactivity versus bead-based multiplex assays. These findings support AADT-PEA as a promising diagnostic and research tool for monitoring complement activation in renal disorders; larger, independent cohorts are warranted for clinical validation
