Purification, Immunological Identification, and Characterization of the Novel Silkworm Pupae Allergen <i>Bombyx mori</i> Lipoprotein 3 (Bomb m 6)
Wenqi Yue, Songyuan Huang, Shiwen Lin, Kan He, Weiyi He, Jiamin Chen, Liuying Li, Wenxiang Chai, Xuli Wu
Abstract
Allergic reactions caused by silkworm pupae greatly limit their utilization, and studies suggest that silkworm pupae proteins of 25–30 kDa may be the principal allergens. To further understand these allergens, we attempted to purify a protein of about 30 kDa by ammonium sulfate salting, pH-graded precipitation, and ion-exchange chromatography. The protein was identified by mass spectrometry and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), western blot, enzyme-linked immunosorbent assays, circular dichroism, and fluorescence spectroscopy analyses. We identified the purified protein as Bombyx mori lipoprotein 3 (Bmlp3), which has high IgE reactivity and is a novel uncharacterized allergen that we named Bomb m 6 according to the WHO/IUIS Allergen Nomenclature Sub-Committee. This allergen is stable against heat, acids, bases, and digestion. In conclusion, we successfully purified and characterized a novel silkworm pupa allergen, which may inform the diagnosis and treatment of silkworm pupa allergies.