[A new counting method of airborne Japanese cedar (Cryptomeria japonica) pollen allergens by immunoblotting using anti-Cry j I monoclonal antibody]

We produced monoclonal antibodies against the major allergen of Japanese cedar (Cryptomeria japonica) pollen, Cry j I. KW-S10 antibody reacted only with Japanese cedar pollen and KW-S91 antibody reacted to most angiospermae pollens as well as Japanese cedar pollen. Using these antibodies, we devised a new counting method of Japanese cedar pollen allergen particles by an immunoblotting technique. Airborne pollen allergens were collected on vaseline coated glass slides or Burkard's sampling tape and were transferred onto polyvinylidene difluoride (PVDF) membranes. The membranes were treated with anti-Cry j I monoclonal antibody conjugated with alkaline phosphatase. Pollen allergens were detected as spots on the membranes after staining with phosphatase substrate (BCIP/NBT). This method using KW-S10 antibody measured only the amounts of allergen from Japanese cedar pollen, while with KW-S91 antibody, the method measured the amounts of pollens which have antigenicity in common with Japanese cedar pollen.