PNP and ALG1 vs. healthy controls

Paraneoplastic pemphigus is an autoimmune mucocutaneous blistering disease. It is caused by an autoimmune reaction in patients with a malignant neoplasms. We here used proteome arrays to perform a comprehensive study of autoimmune targets in paraneoplastic pemphigus, anti-p200 pemphigoid (Ap200P), and healthy controls. Solid phase protein arrays featuring more than 9000 full-length human proteins (ProtoArray v5.1, PAH05251020, ThermoFisher, Waltham, Mass) were screened with diluted 1:2000 sera from patients affected by paraneoplastic pemphigus, Ap200P, and healthy controls. Formed immunocomplexes were detected with a secondary antibody fluorescent signal (Alexa Fluor 647 goat anti-human IgG, A21445, ThermoFisher). Manufacturer’s protocol was followed using recommended reagents: ProtoArray Blocking Buffer Kit (PA055, ThermoFisher), Dylight 550 goat anti-GST (#DY550011-13-001), Cayman chemicals, Ann Arbor, Mich). Microarrays were scanned using the LuxScan HT24 (BioCapital) scanner.