Malaysian cockle (Anadara granosa) XXXX : Identification of IgE-binding proteins and effects of different cooking methods.

@entity1 @entity566 @entity2975 @entity56 @entity369 @entity214
The purpose of this study was to evaluate the effect of different cooking methods on the allergenicity of cockle and to identify proteins most frequently bound by IgE antibodies using a proteomics approach. Raw, boiled, fried and roasted extracts of the cockle were prepared. The protein profiles of the extracts were obtained by separation using @entity214 - @entity2975 gel electrophoresis ( @entity214 -PAGE) and 2-dimensional gel electrophoresis (2-DE). IgE-immunoblotting was then performed with the use of individual sera from @entity1 with @entity566 and the major IgE-binding proteins were analyzed by mass-spectrometry. @entity214 -PAGE of raw extract showed 13 protein bands. Smaller numbers of protein bands were detected in the boiled, fried and roasted extracts. The 2-DE gel profile of the raw extract further separated the protein bands to ~50 protein spots with molecular masses between 13 to 180 kDa and isoelectric point (pI) values ranging from 3 to 10. Immunoblotting of raw extract exhibited 11 IgE-binding proteins with two proteins of 36 and 40 kDa as the major IgE-binding proteins, while the boiled extract revealed 3 IgE-binding proteins. Fried and roasted extracts only showed a single IgE-binding protein at 36 kDa. 2-DE immunoblotting of raw extract demonstrated 5 to 20 IgE reactive spots. Mass spectrometry analysis led to identification of 2 important allergens, tropomyosin (36 kDa) and @entity369 kinase (40 kDa). Heated extracts showed a reduction in the number of IgE-reactive bands compared with raw extract, which suggest that thermal treatment can be used as a tool in attempting to reduce cockle allergenicity. The degree of allergenicity of cockle was demonstrated in the order raw > boiled > fried     roasted. Two important allergens reacting with more than 50% of @entity1 ' sera identified using mass spectrometric approaches were tropomyosin and @entity369 kinase. Thus, allergens found in this study would help in component based diagnosis, management of @entity56 @entity1 and to the standardisation of allergenic test products as tools in molecular allergology.
