# US 2016/0354534 A1 · Apparatus for the extracorporeal treatment of blood > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2016/0354534 A1 - Document ID: US20160354534A1 - Status: Published application - Kind: A1 - Application number: 15/242,775 - Filed: 2016-08-22 - Published: 2016-12-08 - Earliest claimed: 2013-12-17 - Assignees: University of Limerick - Applicants: University of Limerick - Inventors: Jakki Cooney; Todd Fumio Kagawa; Edmond Magner - Web page: https://patentgrep.com/patent/US20160354534A1 ## CPC - A61M 1/3687: {Chemical treatment (A61M1/3675 takes precedence)} - A61L 33/0047: {Enzymes, e.g. urokinase, streptokinase} - A61M 1/3496: {Plasmapheresis; Leucopheresis; Lymphopheresis (A61M1/3472 takes precedence)} - A61M 1/3689: {by biological cells} - C12N 9/52: derived from bacteria {or Archaea} - C12N 11/14: Enzymes or microbial cells immobilised on or in an inorganic carrier - C12Y 304/2111: C5a peptidase (3.4.21.110) - A61M: DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY (introducing media into or onto the bodies of animals A61D7/00; means for inserting tampons A61F13/26; devices for administering food or medicines orally A61J; containers for collecting, storing or administering blood or medical fluids A61J1/05); DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY (surgery A61B; chemical aspects of surgical articles A61L); DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR {(Electrotherapy, e.g. producing anaesthesia by the use of alternating or intermittent currents A61N1/36021)} - A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Y: ENZYMES ## Abstract An apparatus for the extracorporeal treatment of blood comprising an extracorporeal blood circuit (2), a pump (6) configured to provide fluid displacement within the extracorporeal blood circuit, and a reaction chamber (8) connected to the extracorporeal blood circuit and configured to receive blood or plasma from the circuit and treat the blood or plasma. The reaction chamber comprises a protease enzyme immobilized to a support, in which the protease enzyme is specific for, and capable of irreversibly cleaving, a human C5a present in the blood or plasma, wherein the abundance of the human C5a in the treated blood or plasma is less than that in the untreated blood or plasma. The apparatus finds utility in the extracorporeal treatment of blood from patients with inflammatory conditions, especially auto-immune disease and sepsis. ## Claims ### Claim 1 A method for the extracorporeal treatment of blood, the method comprising removing blood or a blood fraction from a patient and reacting the blood or blood fraction with a protease enzyme that is specific for and capable of irreversibly cleaving functional C5a, thereby reducing an abundance of functional C5a in the blood or blood fraction, wherein the protease enzyme is immobilised to a support. ### Claim 2 (depends on 1) The method of claim 1, wherein the protease enzyme is a recombinant bacterial C5a protease comprising SEQ ID NO. 3, or a functional variant thereof having at least 90% sequence identity with SEQ ID NO: 3. ### Claim 3 (depends on 2) The method of claim 2, wherein the functional variant of SEQ ID NO: 3 is SEQ ID NO: 4 or SEQ ID NO: 5. ### Claim 4 (depends on 1) The method of claim 1, wherein the reacting step is carried out with an apparatus comprising: an extracorporeal blood circuit; a pump configured to provide fluid displacement with the extracorporeal blood circuit; and a reaction chamber connected to the extracorporeal blood circuit, the reaction chamber configured to receive the blood or blood fraction from the extracorporeal blood circuit and to treat the blood or blood fraction, wherein the reaction chamber contains the protease enzyme immobilised to the support. ### Claim 5 (depends on 4) The method of claim 4, wherein the protease enzyme is a recombinant bacterial C5a protease comprising SEQ ID NO. 3, or a functional variant thereof having at least 90% sequence identity with SEQ ID NO: 3. ### Claim 6 (depends on 5) The method of claim 5, wherein the functional variant of SEQ ID NO: 3 is SEQ ID NO: 4 or SEQ ID NO: 5. ### Claim 7 (depends on 4) The method of claim 4, wherein the apparatus further comprises separating means adapted to separate the blood or blood fraction into a C5a-containing fraction and a non-C5a containing fraction, wherein the reaction chamber receives the C5a-containing fraction. ### Claim 8 (depends on 7) The method of claim 7, wherein the protease enzyme is a recombinant bacterial C5a protease comprising SEQ ID NO. 3, or a functional variant thereof having at least 90% sequence identity with SEQ ID NO: 3. ### Claim 9 (depends on 8) The method of claim 8, wherein the functional variant of SEQ ID NO: 3 is SEQ ID NO: 4 or SEQ ID NO: 5. ### Claim 10 (depends on 7) The method of claim 7, wherein the apparatus further comprises means configured to recombine the treated C5a-containing fraction with the non-C5a containing fraction. ### Claim 11 (depends on 1) The method of claim 1, wherein the support includes a coordinated transition metal ion and one or more functional groups; the C-terminus of the protease enzyme comprises a first tag and a second tag located distally of the first tag, the second tag being separated from the first tag by a spacer; the first tag comprises a motif capable of covalently reacting with the one or more functional groups; and the second tag comprises a motif capable of interacting with the coordinated transition metal ion. ### Claim 12 (depends on 11) The method of claim 11, wherein the first tag is poly-lysine, poly-glutamate, or poly-cysteine, and the second tag is poly-histidine. ### Claim 13 (depends on 12) The method of claim 12, wherein the protease enzyme is a recombinant bacterial C5a protease comprising SEQ ID NO. 3, SEQ ID NO: 4, or SEQ ID NO: 5. ### Claim 14 (depends on 1) The method of claim 1, wherein the support includes a silica material, a methacrylate, a polyacrylamide, a polypyrrole, or a polysaccharide. ### Claim 15 (depends on 14) The method of claim 14, wherein the silica material is selected from the group consisting of mesoporous silica, a monodispersed mesoporous silicate, and a Ni2+-modified mesoporous silica. ### Claim 16 (depends on 1) The method of claim 1, wherein the support comprises a multiplicity of beads and the protease enzyme is irreversibly immobilized to a surface of the beads. ### Claim 17 A method for treating sepsis, the method comprising removing blood or a blood fraction from a patient having sepsis and reacting the blood or blood fraction with a protease enzyme that is specific for and capable of irreversibly cleaving functional C5a, thereby reducing an abundance of functional C5a in the blood or blood fraction, wherein the protease enzyme is immobilised to a support. ### Claim 18 (depends on 17) The method of claim 17, wherein the protease enzyme is a recombinant bacterial C5a protease comprising SEQ ID NO. 3, SEQ ID NO: 4, or SEQ ID NO: 5. ### Claim 19 A method for treating an inflammatory condition in a human, the method comprising removing blood or a blood fraction from a patient having an inflammatory condition and reacting the blood or blood fraction with a protease enzyme that is specific for and capable of irreversibly cleaving functional C5a, thereby reducing an abundance of functional C5a in the blood or blood fraction, wherein the protease enzyme is immobilised to a support. ### Claim 20 (depends on 19) The method of claim 19, wherein the protease enzyme is a recombinant bacterial C5a protease comprising SEQ ID NO. 3, SEQ ID NO: 4, or SEQ ID NO: 5.