Preparation of cashew patches and application to mice.
Epicutaneous patches were loaded with 50 µg of
cashew protein extract conjugated
to Fluoroprobe-647 (Interchim), prepared in phosphate buffer 0.1 M,
or 50 µl of phosphate buffer 0.1 M
alone (excipient patches). Patches were dried at 30°C in a ventilated
oven and stored at 4°C. Before patch application, mice were
anaesthetized with ketamine and xylazine (50 and 10 mg/kg, respectively)
and hair on the back was removed using electric clippers and depilatory
cream (Reckitt Benckiser). Patches were applied the following day and
secured using an Urgoderm® bandage (Urgo
Laboratories). Patches were maintained for 6 hours based on optimal
timepoints previously defined.