HIGH-LEVEL EXPRESSION OF TREE POLLEN ISOALLERGENS IN ESCHERICHIA-COLI

cDNAs coding for the major allergen of alder (Alnus glutinosa) pollen Aln g 1, for nine isoforms of Bet v 1, the major birch (Betula verruco sa) pollen allergen, and for four isoforms of Cor a 1, the major aller gen of hazel (Corylus avellana) pollen, were inserted into the plasmid pMW175 or pMW 172 and expressed in Escherichia coli as recombinant no n-fusion proteins. These constructs produced between 20 and 160 mg pro tein/l. The recombinant tree pollen isoallergens were tested in immuno blots for their antibody binding properties. For this purpose, we used two monoclonal antibodies (BIP 1 and BIP 4) raised against natural Be t v 1, a polyclonal rabbit anti-recombinant Bet v 1a, as well as serum IgE from allergic patients. Our results show that this expression sys tem is suitable for the production of milligram amounts of tree pollen isoallergens which can be used for the characterization of allergenic epitopes recognized by T and B cells.