Cloning and characterization of Blo t 11, a high molecular weight Blomia tropicalis allergen homologous to paramyosin

Abstract

Three polymorphic variants of Blo t 11 were isolated and characterized. Recombinant Blo t 11 significantly reacted to 52% of IgE from allergic patients. Although multiple IgE and IgG epitopes were mapped throughout the molecule, an immunodominant region (fD) was delineated and characterized. The native Blo t 11, isolated by immunoaffinity purification and confirmed by MALDI-TOF MS, was shown to exist in at least 10 isoforms. Comparative in vitro and in vivo studies between the native and recombinant Blo t 11 showed that fD has comparable IgE reactivity with its native counterpart. Blo t 11 monoclonal antibodies, generated by DNA immunization, were exploited fully for detection, quantitation and isolation of native and recombinant Blo t 11. Furthermore, four Blo t 11 hybridoma clones with mutations on immunoglobulin constant region were characterized.