[An enzyme-linked immunosorbent assay for the quantitation of the major allergen from Japanese cedar (Cryptomeria japonica) pollen, Cry j I, using monoclonal antibodies].
We have developed an enzyme-linked immunosorbent assay (ELISA) using two anti-Cry j I monoclonal antibodies (KW-S10 and KW-S131) for the quantitation of the major allergen from Japanese cedar (Cryptomeria Japonica) pollen, Cry j I. Polystyrene microplates coated with KW-S131 were incubated with pollen allergen extracts. Cry j I, which bound to the antibody, was detected with alkaline phosphatase-conjugated KW-S10 using chromogenic enzyme substrate. Cry j I could be measured in concentration of between 0.16 and 2.5 ng/ml by this assay. Intra- and inter-assay coefficients of variation were 1.1-3.5% and 0.9-4.6%, respectively. This assay was considered that it was specific for Cry j I of Japanese cedar pollen because it didn't react with allergens of hinoki (Chamaecypairs obtusa) pollen which have antigenicity in common to Japanese cedar pollen. This assay would be useful for the standardization of Japanese cedar pollen allergen extracts.
Watanabe
M
M
Kowa Research Institute, Kowa Co., Ltd.
Tamura
M
M
Nagoya
T
T
Takahashi
Y
Y
Katagiri
S
S
Oka
T
T
D000485:Allergens / Q000032:analysis*; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D010940:Plant Proteins; D011058:Pollen / Q000276:immunology*; D014197:Trees
D000485:Allergens; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; C065290:Cry j I protein, Cryptomeria japonica; D010940:Plant Proteins