Lysis/resealing process and device for incorporating an active ingredient, in particular asparaginase or inositol hexaphosphate, in erythrocytes

A lysis/resealing process for preparing erythrocytes which contain an active ingredient, the process comprising the following steps:

• (1) placing a globular erythrocyte concentrate in suspension in an isotonic solution at a haematocrit level which is equal to or greater than 65%, with refrigeration at from +1 to +8° C., thereby obtaining an erythrocyte suspension;
• (2) measuring the osmotic fragility based on a sample of erythrocytes from the globular erythrocyte concentrate, the steps 1 and 2 being able to be carried out in any order;
• (3) lysing the erythrocytes in the erythrocyte suspension inside a first chamber, at a temperature which is constantly maintained at from +1 to +8° C., by allowing the erythrocyte suspension obtained in step (1) and a hypotonic lysis solution which is refrigerated at from +1 to +8° C., to circulate in a dialysis cartridge, thus obtaining a suspension of lysed erythrocytes, wherein, based on the measurement of osmotic fragility, the flow rate of the erythrocyte suspension which passes into the dialysis cartridge is adjusted, or the osmolarity of the lysis solution is selected;
• (4) internalizing an active ingredient in the erythrocytes by adding the active ingredient to the erythrocyte suspension obtained in step (1) or in step (3) before or after passing the erythrocyte suspension into the dialysis cartridge, thereby obtaining internalization of the active ingredient in the erythrocytes;
• and
• (5) resealing the erythrocytes obtained at step (4) in a second chamber at a temperature of from +30 to +40° C. by means of a hypertonic solution.