Fig. 7: Patient-derived antibody production and antigen discovery. | Nature Communications

Fig. 7: Patient-derived antibody production and antigen discovery.

From: B cell profiles, antibody repertoire and reactivity reveal dysregulated responses with autoimmune features in melanoma

Fig. 7: Patient-derived antibody production and antigen discovery.The alternative text for this image may have been generated using AI.

a Schematic explaining patient-derived antibody heavy and light chain cloning and production of recombinant human IgG1. b Coomassie SDS-PAGE of monoclonal antibodies bearing patient-derived variable region sequences in reducing and non-reducing conditions, annotated with kappa/lambda information. c Schematic of immuno-mass spectrometry experimental procedure and analysis pipeline for antigen discovery. Left panel: pull down of normal skin protein lysate or human melanoma protein lysate using patient-derived IgG1, anti-CSPG4 IgG1 (positive control) and anti-NIP IgG1 (negative control), followed by trypsin digestion and Shotgun Mass Spectrometry LC-MS/MS; Right panel: Shotgun Mass-Spectrometry LC-MS/MS output filtering criteria to select potential antigen peptides of interest to be confirmed by Parallel Reaction Monitored Mass Spectrometry (PRM-MS). d Summary of the immuno-mass spectrometry (blue boxes show the number of peptides pulled down by the antibodies from melanoma tissue protein lysate; red outlines represent an antibody reaction with a specific protein from normal skin protein lysate as well) and glycan array (yellow boxes show antibody reaction with specific glycans) screening results for antigen discovery with patient-derived antibodies. Source data are provided as a Source data file.

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