# US 10,307,510 B2 · Methods of removing alpha-galactose > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,307,510 B2 - Document ID: US10307510B2 - Status: Granted - Kind: B2 - Application number: 14/532,089 - Filed: 2014-11-04 - Published: 2015-05-07 - Granted: 2019-06-04 - Earliest claimed: 2013-11-04 - Assignees: LifeCell Corporation - Applicants: LifeCell Corporation - Inventors: Hui Xu; Li Ting Huang; Hua Wan; Rick T. Owens; Nathaniel Bachrach - Web page: https://patentgrep.com/patent/US10307510B2 ## CPC - A61L 27/3687: {characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents} - A61L 27/24: Collagen - A61L 27/362: {Skin, e.g. dermal papillae} - A61L 27/3695: {characterised by the function or physical properties of the final product, where no specific conditions are defined to achieve this (A61L27/3687, A61L27/3691 take precedence)} - A61L 27/38: {containing added animal cells (organs or tissue containing native cells A61L27/36)} - A61L 27/54: Biologically active materials, e.g. therapeutic substances {(A61L27/227 takes precedence)} - A61L 27/58: Materials at least partially resorbable by the body - G01N 33/573: for enzymes or isoenzymes - A61L 2300/412: Tissue-regenerating or healing or proliferative agents - A61L 2430/34: for soft tissue reconstruction - A61L 2430/40: Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking - 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) - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract Tissue products lacking a desired percentage of immunogenic epitopes, such as galactose alpha-1,3 galactose epitopes, are provided. Methods of making and using the tissue products are also provided. ## Claims ### Claim 1 A method of preparing a dermal tissue product, comprising: selecting at least one collagen-containing tissue matrix contained within a cellular dermal tissue and containing galactose alpha-1,3-galactose moieties; and contacting the at least one collagen-containing tissue matrix contained within the cellular dermal tissue with alcalase under conditions sufficient to remove galactose alpha-1,3-galactose moieties from the tissue matrix contained within the cellular dermal tissue; thereby preparing a dermal tissue product. ### Claim 2 (depends on 1) The method of claim 1, wherein the method further comprises decellularizing the cellular dermal tissue following contacting the at least one collagen-containing tissue matrix contained within the cellular dermal tissue with alcalase. ### Claim 3 (depends on 2) The method of claim 2, wherein the tissue matrix is completely decellularized. ### Claim 4 (depends on 1) The method of claim 1, wherein galactose alpha-1,3-galactose is substantially or completely removed from the collagen-containing tissue matrix. ### Claim 5 (depends on 1) The method of claim 1, wherein the alcalase removes galactose alpha-1,3-galactose moieties without damaging the at least one tissue matrix. ### Claim 6 (depends on 1) The method of claim 1, further comprising performing an assay to determine if galactose alpha-1,3-galactose moieties have been removed from the at least one collagen-containing tissue matrix. ### Claim 7 (depends on 6) The method of claim 6, wherein the assay comprises measuring a concentration of galactose alpha-1,3-galactose moieties in the collagen-containing tissue matrix. ### Claim 8 (depends on 6) The method of claim 6, wherein the assay comprises an histochemical assay. ### Claim 9 (depends on 6) The method of claim 6, wherein the assay comprises an immunoassay. ### Claim 10 (depends on 1) The method of claim 1, wherein the alcalase is used at a concentration ranging from about 0.0001% to about 0.1% and for a period of time ranging from about 0.5 hours to about 24 hours. ### Claim 11 (depends on 1) The method of claim 1, further comprising contacting the tissue with alpha-galactosidase following contacting the at least one collagen-containing tissue matrix contained within the cellular dermal tissue with alcalase. ### Claim 12 (depends on 1) The method of claim 1, wherein tissue from one or more different animals are used. ### Claim 13 (depends on 1) The method of claim 1, wherein the at least one collagen-containing tissue matrix is a porcine tissue matrix. ### Claim 14 (depends on 2) The method of claim 2, further comprising adding one or more viable and histocompatible cells to the tissue product. ### Claim 15 (depends on 14) The method of claim 14, wherein the one or more cells are mammalian cells. ### Claim 16 (depends on 14) The method of claim 14, wherein the one or more cells are stem cells. ### Claim 17 (depends on 1) The method of claim 1, further comprising treating the at least one collagen-containing tissue matrix to reduce bioburden. ### Claim 18 (depends on 17) The method of claim 17, wherein treating the at least one collagen-containing tissue matrix to reduce bioburden comprises irradiating the tissue product.