Treatment of High Molecular Mass Antigen of Paracoccidioides brasiliensis with Papain and Detection of Antigenic Subcomponents
Keywords:
Fungus, Immunodiagnosis, Paracoccidioidomycosis, VaccineAbstract
Paracoccidioidomycosis (PCM) is a chronic systemic granulomatous disease caused by the dimorphic fungi of Paracoccidioides genus, which produce high molecular weight (hMM) antigens that induce a protective Th1 immune response. The smaller but still antigenic/immunogenic fragments may be useful for vaccine purposes as well as for immunodiagnosis. This study aimed to partially characterize components of the hMM fraction by enzymatic treatment with papain. Initially, the hMM fraction was obtained by chromatography from cell-free antigen produced by P. brasiliensis strain 18 (Pb18). The fraction was then treated with papain at 37°C for 4 h and subsequently fractionated by High Performance Liquid Chromatography (HPLC). The resulting fractions were analyzed initially by dot blotting and then by Western blotting, using IgG from a pool of sera of patients with PCM. The results revealed the positivity of two fractions in the dot blotting and the western blotting indicated the presence of a high molecular mass band and another of approximately 70kDa. We conclude from these preliminary data that the treatment of the hMM fraction with the enzyme papain can generate a subcomponent of approximately 70kDa with antigenic activity.
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Copyright (c) 2025 Mari Sumigawa Kaminami, Bianca Dorana de Oliveira Souza, Audrey de Souza Marquez, Adriane Lenhard-Vidal, Flávio Hiroshi Itano, Franciele Ayumi Semencio Chiyoda-Rodini, Igor Massahiro de Souza Suguiura, Mario Augusto Ono, Eiko Nakagawa Itano

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