The Use of Combinatorial Hexapeptide Ligand Library (CPLL) in Allergomics
The recent progress of proteomic protocols led to more efficient protein extraction and concentration procedures to remove nonprotein interfering compounds present in the starting material and to increase the concentration of underrepresented proteins. Combinatorial hexapeptide ligand libraries (CPLL) were recently applied to both plant- and animal-derived tissues for capturing the low- and very low-abundance allergens. Several IgE-binding proteins which were previously absent or poorly represented by using conventional proteomics tools have been detected and characterized through a CPLL-based approach. In the present chapter, a protocol based on improved protein extraction and enrichment by CPLL, allowing the immunochemical characterization of several “hidden allergens” in cypress pollen, is described in detail.
Key wordsHexapeptide ligand libraries Low-abundance allergens Pollen allergens Mass spectrometry Proteomics
Collaboration with Drs. Egisto Boschetti and Pier Giorgio Righetti, the pioneer developers of the CPLL technology, is gratefully acknowledged and was at the basis of this protocol and its adaptation to allergomics studies.
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