IMMUNOBIOCHEMICAL CHARACTERIZATION OF BRASSICA-CAMPESTRIS POLLEN ALLERGEN

Brassica campestris (BC), Eng. Mustard, is an important source of poll en allergen, responsible for type I hypersensitivity disorders. In the present study, BC pollen extract was characterized by TLIEF, SDS-PAGE and immunoprinting. The extract separated into 50 silver stained band s of pI 3-9 on isoelectric focusing whereas it resolved into 14 Coomas sie blue stained protein bands of 14-100 kD on SDS-PAGE. Immunoblot an alysis with individual patient sera detected four allergenic proteins of 90, 67, 60 and 14 kD. BC separated into 8 peaks (Bras 1-8) on DEAE Sephadex A-50 column. Bras 2 was found to be most potent by IgE specif ic ELISA, hence further fractionated on Sephadex G-200. A protein of 9 0 kD (Bras 2a) isolated by gel filtration was found to be most allerge nic protein by ELISA inhibition. The findings shall be applicable in s tandardization of future batches of BC pollen extract to be used for a llergy diagnosis and immunotherapy.