Characterization of a monoclonal antibody (P40) against the 68 kD major allergen ofPenicillium notatum

摘要

SummaryA monoclonal antibody (MoAb P40) against the 68 kD major allergen ofpenicillium notatum (P. notatum)was obtained by immunizing the mouse with a crude extract ofP. notatum.Analysed by two‐dimensional gel electrophoresis and immunoblotting, P40 reacted with two different isoforms of the 68 kD component ofP. notatumwith pIs of 5.4 and 5.5. In addition toP. notatum, P40 showed positive ELISA activity toAspergillus fumigatus (A. fumigatus) but not to components of six other fungi includingAlternaria porri, Cladosporium cladosporoides, Aureobasidium pullulans, Fusarium solani, Rhizopus arrhizusandCandida albicans.Analysed by ELISA, MoAb P40 also showed positive activity to two (P. frequentansandP. roseopurpureum) of the 10 otherPenicilliumspecies and two (A. terreusandA. flavus) of the four otherAspergillusspecies tested. SDS‐PAGE and immunoblotting studies demonstrated P40 positive reactivity to components with MW of about 67 kD in all thesePenicilliumandAspergillusspecies with positive ELISA activity to P40. Furthermore, immunoblotting activity of MoAb P40 to the 67 kD component ofA. nigerwas also observed. The epitope of the 68 kD allergen ofP. notatumrecognized by MoAb P40 was resistant to treatment of periodate oxidation with concentration of NaIO4up to 20 mm. This MoAb may thus be useful in the characterization and purification of the 68 kD allergen from crude extracts, and in the molecular cloning of allergen ge