# US 2014/0154797 A1 · Lysis/Resealing Process and Device for Incorporating an Active Ingredient, in Particular Asparaginase or Inositol Hexaphosphate, in Erythrocytes > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0154797 A1 - Document ID: US20140154797A1 - Status: Published application - Kind: A1 - Application number: 14/089,834 - Filed: 2013-11-26 - Published: 2014-06-05 - Earliest claimed: 2004-08-05 - Assignees: Erytech Pharma - Applicants: Erytech Pharma - Inventors: Yann GODFRIN - Web page: https://patentgrep.com/patent/US20140154797A1 ## CPC - C12M 47/06: {Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material (lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003)} - A61K 9/5068: {Cell membranes or bacterial membranes enclosing drugs (liposomes with additional exogenous lipids A61K9/127)} - A61K 31/6615: Compounds having two or more esterified phosphorus acid groups, e.g. inositol triphosphate, phytic acid - A61K 35/18: Erythrocytes (haemoglobin A61K38/42) - A61P 7/00: Drugs for disorders of the blood or the extracellular fluid - A61P 35/00: Antineoplastic agents - A61P 35/02: specific for leukemia - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract A lysis/resealing process for preparing erythrocytes containing active ingredient is provided comprising placing a globular concentrate in suspension in an isotonic solution having a haematocrit level which is equal to or greater than 65%, with refrigeration at 1 to 8° C.; measuring the osmotic fragility based on a sample of erythrocytes from that same globular concentrate, preferably on a sample of the suspension; lysis and internalisation procedure of the active ingredient, inside the same chamber, at a temperature maintained at 1 to 8° C., comprising allowing the erythrocyte suspension having a haematocrit level equal to or greater than 65% and a hypotonic lysis solution which is refrigerated at 1 to 8° C., to circulate in a dialysis cartridge; the lysis parameters being adjusted in accordance with the osmotic fragility previously measured; and resealing in a second chamber at a temperature of from 30 to 40° C. by means of a hypertonic solution. ## Claims ### Claim 1 A lysis device for preparing erythrocytes which contain an active ingredient, wherein the device comprises a module which may be cooled, and a sterile, single-use removable assembly which is configured in order to be able to be placed in the module and which comprises a dialysis cartridge which can be connected, at one side, to an inlet for a lysis solution and, at the other side, to an inlet for an erythrocyte suspension, wherein the module maintains the temperature at from +1 to +8° C. and adjusts the flow rate of the erythrocyte suspension through the dialysis cartridge and/or adjusts the osmolarity of the lysis solution, in accordance with the osmotic fragility of the erythrocytes to be processed. ### Claim 2 (depends on 1) The device according to claim 1, wherein the removable assembly comprises a pouch which can contain the erythrocyte suspension and a tube which connects that pouch to the dialysis cartridge, and the module comprises a pump which can co-operate with that tube and cause the erythrocyte suspension to circulate from the pouch towards and through the cartridge. ### Claim 3 (depends on 2) The device according to claim 2, wherein the pouch is provided with a loop type tube which is connected to the pouch at the two ends thereof, and the module comprises a pump which can co-operate with that tube and bring about circulation of the contents of the pouch to and from that pouch. ### Claim 4 (depends on 2) The device according to claim 2, wherein a tube for injecting the active ingredient is connected to the tube which connects the pouch to the blood inlet of the dialysis cartridge, the cartridge is connected by a tube to a flask which can contain the lysis solution, and the module is configured to receive said flask and a pump which can co-operate with the tube in order to cause the lysis solution to circulate towards and through the dialysis cartridge; and the blood outlet of the dialysis cartridge is connected to a tube which opens, or which may open, outside the module. ### Claim 5 (depends on 4) The device according to claim 4, wherein the tube which opens, or which may open, outside the module is connected to a second pouch which can collect the erythrocyte suspension which is discharged from the lysis, as well as a resealing solution, that pouch being arranged in a second module which can control the temperature in the module at from +30 to +40° C. ### Claim 6 (depends on 2) The device according to claim 2, wherein said pump which causes the erythrocyte suspension to circulate from the pouch towards and through the cartridge is connected to the module that adjusts the flow rate. ### Claim 7 (depends on 2) The device according to claim 2, wherein a tube for injecting the active ingredient is connected to the tube which connects the pouch to the <> inlet of the dialysis cartridge. ### Claim 8 (depends on 2) The device according to claim 2, wherein the dialysis cartridge is connected by a tube to a flask which can contain the lysis solution, and the refrigerated module can receive that flask and a pump which can co-operate with the tube in order to cause the lysis solution to circulate towards and through the dialysis cartridge. ### Claim 9 (depends on 1) The device according to claim 1, wherein the module is capable of maintaining a temperature of from +2 to +6° C. ### Claim 10 (depends on 2) The device according to claim 2, wherein the blood outlet of the dialysis cartridge is connected to an outlet tube which opens, or which may open, outside the module and a tube for injecting the active ingredient is connected to that outlet tube. ### Claim 11 (depends on 1) The device according to claim 1, wherein the module is configured to control the pump which conveys the erythrocyte suspension towards the dialysis cartridge. ### Claim 12 (depends on 1) The device according to claim 1, wherein the module is configured to control the osmolarity of a lysis solution, either in order to dilute and lower the osmolarity, or to increase that osmolarity by a suitable solute being introduced. ### Claim 13 (depends on 1) The device according to claim 1, wherein the lysis process may be controlled electronically and wherein optionally the resealing process may be controlled electronically, and wherein both processes may be controlled in accordance with instructions which are input by the operator, or in accordance with data input by the operator, with reference to the osmotic fragility. ### Claim 14 (depends on 13) The device according to claim 13, further comprising temperature sensors which allow the temperature in the modules and/or at the temperature sensor for the erythrocyte suspension to be controlled. ### Claim 15 (depends on 1) The device according to claim 1, wherein said active ingredient is asparaginase. ### Claim 16 (depends on 1) The device according to claim 1, wherein said active ingredient is inositol phosphate.