Supplementary legends to the figures
Figure S1: Analysis of on-site prepared cashew protein extracts
by SDS PAGE. Protein fraction was extracted from defatted cashew flour
as indicated in material and methods. This extract was denatured and
analyzed by SDS PAGE (lane 2) in comparison to initial cashew flour
(lane 1) and commercial cashew protein extract (Stallergenes Greer, lane
3). The relative molecular mass (kDa) is indicated on the left. The
position of major allergens are indicated on the right, based on
previous published data . Note that Ana o 2 separates as two distinct
bands following denaturation since it is constituted by two subunits
(acidic and basic) linked by disulfide bonds.
Figure S2: Analysis of cashew protein extracts recovered from
patches. Epicutaneous patches were loaded with commercial cashew
protein extracts (Stallergenes Greer) in liquid form, by dissolving
protein lyophilizate in PBS 1X, NaCl 0.45% or phosphate buffer 0.1 M.
Patches were dried for one hour at 30°C. Then, proteins were redissolved
from patch backing using distilled water. (A ) Recovered
proteins were used as a coating in ELISA to analyze their capacity to
bind to cashew-specific antibodies induced in mice by oral
sensitization. HRP-conjugated anti-mouse IgG1 (left panel) or anti-mouse
IgE (right panel) were used as secondary antibodies. Optical densities
(O.D.) at 450 nm were plotted against plasma dilution, (n = 7 per
group). Data are median and range of individual values and non-linear
regression curve was obtained using sigmoidal 4PL equation. (B )
Recovered protein extracts were centrifuged and total extract (before
centrifugation, T), supernatant (S) and pellet (P) were denatured and
analyzed by SDS PAGE. The relative molecular mass (kDa) is indicated on
the left.