In this study, the objective is the identification of allergens recognized by the IgE antibodies from allergic patients. Genclis SAS used the Precellys® Evolution to homogenize food samples for protein extraction. The quality of proteins extracted with the Precellys® Evolution was compared by Coomassie blue staining and by western blot to two other homogenizers: the IKA Ultra-Turrax® dispenser and the Awel Mixwell® blender.
The protein profiles of the 3 samples were compared by Coomassie blue staining (Figure 1) and by western blot for the cashew nut samples only (Figure 2). The cashew nut allergens were recognized by specific IgE antibodies, as observed by western blotting. Similar protein and allergenicity profiles were obtained with Precellys® Evolution compared to the two other homogenizers compared in this study.
Figure 1: Comparison by Coomassie staining of the food extracts homogenized with the Precellys Evolution® (PE), the IKA Ultra-Turrax® disperser (IK) and the Awel Mixwell® blender (AM). Lane 1 – dog biscuits/PE; 2 – dog biscuits/IK; 3 – dog biscuits/AM; 4 – cashew nuts/PE; 5 – cashew nuts/IK; 6 – cashew nuts/AM; 7 – green beans/PE; 8 – green beans/ IK; 9 – green beans/AM.
Figure 2: IgE reactivity was compared in cashew nut extracts by western blot after homogenization with the 3 instruments. Samples were loaded onto a 12% polyacrylamide gel, and separated by SDS PAGE electrophoresis, followed by transfer to PVDF membrane. The membrane was incubated with the serum of a patient allergic to cashews, followed by probing with a secondary anti-Human IgE HRP-conjugated antibody. Lane 1 – cashew nuts/PE; 2 – cashew nuts/PE; 3 – cashew nuts/IK; 4 – cashew nuts/AM.