High throughput screening: a rapid way to recombinant allergens

Complex allergenic sources such as moulds, foods and mites contain large pa nels of IgE-binding molecules which need to be cloned, produced and charact erized in order to mimic the entire allergenicity of whole extracts reconst ituted by mixing single standardized recombinant allergens. Phage display o f cDNA libraries allows selective enrichment of allergen-expressing clones using IgE from allergic patients. For the characterization of all different clones present in enriched cDNA libraries in a fast and cost-effective way , however, high-throughput screening technology is required. We have used a high-throughput, quantitative technology for fast identification of all di fferent clones present in selectively enriched phage surface-displayed cDNA libraries in order to characterize whole allergenic repertoires from compl ex allergenic sources. The strategy, based on a combination of phage displa y and high-density arrays, allowed fast discovery of panels of related stru ctures from different allergenic sources. They cover secreted, cytoplasmic and structural proteins with or without enzymatic activity and offer a rati onal explanation for the IgE-mediated cross-reactivity frequently encounter ed in clinical practice.