INVESTIGADORES
NUSBLAT Alejandro David
artículos
Título:
Novel antigen as a promising tool for immunodiagnosis of histoplasmosis
Autor/es:
AGUSTINA TOSCANINI; GONZALEZ MAGLIO; PAULA CAPECE; GLADYS POSSE; CRISTINA A. IOVANNITTI; A. NUSBLAT; CUESTAS MARIA LUJAN
Revista:
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Editorial:
SPRINGER
Referencias:
Lugar: Berlin; Año: 2020
ISSN:
0175-7598
Resumen:
Few antigens are currently used aslaboratory tools for the immunodiagnosis of histoplasmosis, the H and Mglycoproteins, the galactomannan antigen and the largely unknown and poorlycharacterized Histoplasma capsulatumpolysaccharide antigen (HPA). The broad application of H and Mantigens for antibody detection is limited by their low sensitivity andspecificity, particularly, in the immunocompromised host, and current methodsfor their production are time-consuming and problematic. Furthermore, Histoplasma antigen detection tools areoften unavailable in resource-poor countries, such as regions of Central andSouth America where histoplasmosis is endemic. The protein of 100 kDa of H. capsulatum (Hcp100) or its codifying gene was proposed as anovel target for histoplasmosis therapy and diagnosis due to its essential roleduring infection. In this work, the codon sequence of Hcp100 was clonedand expressed as a secretory protein in Pichiapastoris. After optimizing the culture conditions and purifying by immobilizedmetal ion affinity chromatography, the highest yield of Hcp100 reachedapproximately 1 mg/L with 90% purity in shake flasks using basal salt mediumsupplemented with casamino acids after 72 h of methanol induction.Immunoreactivity of the purified rHcp100 was assessed.Thisstudy reveals that P. pastoris is anexcellent system for recombinant Hcp100 expression with potential applicationsin diagnosis, prognosis and molecular targeted therapy. Finally, these resultsmay provide an alternative strategy for the production of M and H antigens soas to replace their native forms used in immunodiffusion and immunoblottingassays for diagnosis of histoplasmosis.