In our continuing effort to improve the standardization program of allergenic extracts, our laboratory is actively pursuing the project in the identification and molecular characterization of major allergens. Our studies include: Latex allergans: Two acidic and one basic proteins are most reactive to patient serum. Using Rotophor (separate proteins by IEF) and 2-D gel, single species of proteins can be identified. These proteins will further be purified by HPLC and sequenced. Apple allergans: We have successfully synthesized random peptide library on acrylamide beads using selective peptide technology. Comparing both selective and combinatorial peptide library technology, we obtained the same epitopes which have significant inhibition activity on our monoclonal antibody. Further studies will apply these technologies to determine the epitope from purified allergenic proteins. We are also screening sera which has IgE response to specific protein bands of apple extract. We have found that 13K, 17K, and 31K dalton proteins in the apple extract are reactive to IgE. Currently, the amino terminal sequences of each band are being determined. Obtained amino acid sequences information will be useful for specific allergan cloning. We also collected different growth stages of apple samples from Western Maryland Research and Educations Center of the University of Maryland. Investigation of the nucleic acid and protein levels in response to IgE binding is in the process. Major allergans from tree pollens, molds, and peanuts will also be studied.
