Lab scale and medium scale production of recombinant allergens in Escherichia coli

• Published in: Methods (San Diego, Calif.) Vol. 32; no. 3; pp. 219 - 226
• Main Authors: Wallner, Michael, Gruber, Petra, Radauer, Christian, Maderegger, Bernhard, Susani, Markus, Hoffmann-Sommergruber, Karin, Ferreira, Fátima
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.03.2004
• Subjects: Allergens - biosynthesis; Allergens - genetics; Allergens - isolation & purification; Cloning, Molecular - methods; Escherichia coli; Genetic Vectors
• ISSN: 1046-2023; 1095-9130
• DOI: 10.1016/j.ymeth.2003.08.004

Recombinant products have become invaluable tools for diagnostic as well as therapeutic purposes in modern medicine. Especially in cases where raw naturally derived products are difficult to standardize, well-defined recombinant single components represent the matter of choice. In the recent past, much effort has been undertaken to define individual proteins derived from various sources like pollen, spores of moulds, pet dander, and food causing Type 1 allergic reactions in humans. Therefore, methods for cloning, sequencing, and expressing cDNAs coding for allergens in Escherichia coli became of great interest to allergologists. For the recombinant production of allergens, suitable expression systems, growing conditions, and purification steps have to be established for each individual product. Finally, the purified recombinant allergen has to be carefully investigated for the biochemical, biophysical, and immunological properties. In the following paper, several prokaryotic expression systems, purification strategies, and analytical methods will be presented and pitfalls discussed.