Protein chips for detection of mite allergens using Kunitz‐type protease inhibitors

Abstract

Stored‐food and house‐dust arthropods include many species of mites and beetles that affect human health. For diagnostic tests proteases such as trypsin are utilized as they are indicators of the presence of allergen contaminants in food. We recently characterized Kunitz‐type protease inhibitors (KPIs) from Solanum palustre. Here we studied biotechnological applications of KPI‐B1 and ‐B4. We manufactured a protein chip with immobilized KPI‐B1 and ‐B4 and showed trypsin/chymotrypsin‐binding specificity, indicating that the recombinant proteins have protease selectivity. We employed the protein chip to capture mite proteins belonging to the protease family with polyclonal anti‐mite antibodies. The mite diagnostic chip can be useful for detecting mite allergens.

Environmental assessment and exposure control of dust mites: a practice parameter

“…Assays for dust mite allergen proteins also are available in a multiplex format in combination with allergenic proteins from up to 5 other allergenic species, 46 in a chip format for general mite species present in food, 47 and in a screening format (http://inbio.com/US/Products/Rapid-Test-and-Dust-Collection), although this method has some limitations in that it measures proteins in mites and not just dust mites, including Aleuroglyphus ovatus, A siro, Lepidoglypus destructor, Tyrophagus putrescentiae , and Tribolium castaneum . In addition, the test is in the research phase and currently not commercially available.…”