# US 2015/0147408 A1 · Method validation unit

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2015/0147408 A1
- Document ID: US20150147408A1
- Status: Published application
- Kind: A1
- Application number: 14/404,925
- Filed: 2013-05-30
- Published: 2015-05-28
- Earliest claimed: 2012-05-31
- Assignees: Austrialian Biotechnologies PTY. Limited
- Applicants: Australian Biotechnologies PTY. Limited
- Inventors: Simon Jonothan Berry; Sharon Bryce; Jerome Alon Goldberg
- Web page: https://patentgrep.com/patent/US20150147408A1

## CPC

- C12Q 1/22: Testing for sterility conditions
- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- A61L 2/04: Heat (radiation A61L2/08)
- A61L 2/28: Devices for testing the effectiveness or completeness of sterilisation or disinfection, e.g. indicators which change colour
- G01N 31/226: {for investigating the degree of sterilisation}
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- 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)
- 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

A Method Validation Unit (MVU) for the evaluation of the level of sterilisation in a sterilisation method involving the use of a fluid at or near the supercritical pressure and temperature for that fluid, wherein the MVU comprises a sterilisation indicator housed within a gas-permeable container, wherein the sterilisation indicator comprises an indicator medium and a population of one or more colony forming units (CFUs), and wherein the indicator medium comprises one or more structural features representative of the internal structure of a material to be sterilised in the sterilisation method.

## Claims

### Claim 1

A Method Validation Unit (MVU) for the evaluation of the level of sterilisation in a sterilisation method involving the use of a fluid at or near the supercritical pressure and temperature for that fluid, wherein the MVU comprises:
a sterilisation indicator housed within a gas-permeable container, wherein the sterilisation indicator comprises an indicator medium and a population of one or more colony forming units (CFUs), and wherein the indicator medium comprises one or more structural features representative of the internal structure of a material to be sterilised in the sterilisation method.

### Claim 2 (depends on 1)

A MVU according to claim 1 wherein the population of the one or more CFUs is at least 103 CFUs, 106 CFUs or 1012 CFUs.

### Claim 3 (depends on 1, 2)

A MVU according to claim 1 or 2, wherein the population of the one or more CFUs is 2×106 CFUs.

### Claim 4 (depends on 1, 2, 3)

A MVU according to anyone of claims 1 to 3, wherein the indicator medium comprises a porous structure.

### Claim 5 (depends on 1, 2, 3, 4)

A MVU according to anyone of claims 1 to 4 wherein the gas-permeable container is substantially gas-permeable over its entire surface.

### Claim 6 (depends on 1, 2, 3, 4, 5)

A MVU according to anyone of claims 1 to 5 wherein the container comprises a non-woven polymeric material.

### Claim 7 (depends on 1, 2, 3, 4, 5, 6)

A MVU of any one of claims 1 to 6, wherein the MVU is housed within a second gas-permeable container.

### Claim 8 (depends on 1, 2, 3, 4, 5, 6, 7)

A MVU according to any one of claims 1 to 7 wherein the indicator medium comprises one or more materials selected from: tissue suitable for transplant, research or therapeutical grade bone, demineralised bone, demineralised bone matrix, a paste comprising demineralised bone or demineralised bone matrix, whole cortical bone pieces, tendon, Achilles tendon, cartilage, ligament, skin, connective or musculoskeletal tissue or biological tissue suitable for implantation; natural or synthetic polymer, biomedical polymer, medical-grade polymer, biodegradable polymer, synthetic human tissue analogue; biologically active molecules, pharmaceutically active compounds, pharmaceutical carriers, or pharmaceutical delivery vehicles; metals, alloys, medical equipment, instruments, prosthetics, implants or representative analogues thereof.

### Claim 9 (depends on 1, 2, 3, 4, 5, 6, 7, 8)

A MVU according to any one of claims 1 to 8 wherein the indicator medium comprises Cortical Cancellous Crunch.

### Claim 10 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9)

A MVU according to anyone of claims 1 to 9 wherein the indicator medium comprises human-derived tissue.

### Claim 11 (depends on 8)

A MVU according to claim 8 wherein the synthetic tissue analogue comprises silica, glass or a ceramic material.

### Claim 12 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11)

A MVU of any one of claims 1 to 11 wherein the indicator medium is granular, powder or fibrous in nature and/or formed into a solid mass.

### Claim 13 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)

A MVU of any one of claims 1 to 12 wherein the indicator medium comprises granules of between about 1 and 9 mm, 2 and 7 mm, or 3 and 5 mm, and where the indicator medium comprises a powder, the powder has a mean particle diameter of about 10 to 900 μm, 100 to 700 μm, or 150 to 350 μm.

### Claim 14 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)

A MVU according to anyone of claims 1 to 13 wherein the one or more CFUs are selected from bacteria, vegetative microbial cells, moulds, single-celled organisms, protozoa, yeasts, viruses, or other infective agents spores, or progenitor species thereof.

### Claim 15 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)

A MVU according to anyone of claims 1 to 14 wherein the one or more CFUs are selected from B. Atrophaeus (formerly B. subtilis var Niger), B. stearothermophilus, B. subtilis, B. pumilus, B. cereus, Listeria innocua, Staphylococcus aureus, Salmonella salford, Psuedomonas aeruginosa, Escherichia coli, Preoteus vulgaris, Legionella dunnifii; spores, or progenitor species thereof, or wherein the one or more CFUs is a 2×106 B. Atrophaeus spore suspension.

### Claim 16 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)

A MVU according to any one of claims 1 to 15, wherein the indicator medium has been pre-treated with a SCF, or wherein the indicator medium has been pre-treated with a SCF and wherein that SCF was SCCD.

### Claim 17 (depends on 16)

A MVU according to claim 16 wherein the pre-treatment has removed 25 to 99% of the mass of the material pre-treated, or has removed 70 to 98% of the mass of the material pre-treated, or has removed 85 to 95% of the mass of the material pre-treated.

### Claim 18 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)

A blister pack comprising two or more MVUs according to anyone of claims 1 to 17, wherein the MVUs are detachable from the blister pack.

### Claim 19 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)

A sealable gas-permeable container for use in a sterilisation method, wherein the sealable container comprises a MVU according to anyone of claims 1 to 17.

### Claim 20

A MVU kit for use in the evaluation of a sterilisation method, the kit comprising:
a sealable gas-permeable container, a sterilisation indicator medium, and a carrier medium, wherein the indicator medium comprises one or more structural features representative of the internal structure of the material to be sterilised in the sterilisation method, and wherein the carrier medium comprises a population of one or more CFUs, wherein the indicator medium and the carrier medium are combinable to form a sterilisation indicator.

### Claim 21 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)

A method of manufacturing a MVU according to anyone of claims 1 to 17 comprising the step of housing the sterilisation indicator in the gas-permeable container, wherein the sterilisation indicator comprises an indicator medium and a population of one or more CFUs, and wherein the indicator medium comprises one or more structural features representative of the internal structure of the material, to be sterilised in the sterilisation method, and wherein the MVU is useful in evaluating the level of the sterilisation in the sterilisation method.

### Claim 22 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)

A method of manufacturing a sterilisation indicator as defined in anyone of claims 1 to 17 comprising the steps of: (i) optionally substantially sterilising the indicator medium; (ii) optionally pre-treating the indicator medium with a SCF; (iii) treating the indicator medium with a carrier medium, the carrier medium comprising a population of one or more CFUs, (iv) optionally incubating the treated indicator medium.

### Claim 23 (depends on 22)

A method of manufacturing according to claim 22, wherein the indicator medium is pre-treated with a SCF, or wherein the indicator medium is pre-treated with a SCF and wherein the SCF is SCCD.

### Claim 24 (depends on 22, 23)

A method of manufacturing according to any one of claim 22 or 23 wherein the pre-treatment removes 25 to 99% of the mass of the material pre-treated, or removes 70 to 98% of the mass of the material pre-treated, or removes 85 to 95% of the mass of the material pre-treated.

### Claim 25 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)

Use of a MVU according to any one of claims 1 to 18 in an apparatus for sterilisation, wherein the sterilisation method comprises the use of a fluid at or near the supercritical pressure and temperature for that fluid.

### Claim 26

A sterilisation method comprising bringing a MVU and a material in need of sterilisation into contact with a sterilant fluid, the sterilant fluid comprising a fluid at or near the supercritical pressure and temperature for that fluid, and wherein the MVU is useful in evaluating the level of the sterilisation in the sterilisation method.

### Claim 27 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 26)

A sterilisation method according to claim 26 wherein the MVU is a MVU according to any one of claims 1 to 18.

### Claim 28 (depends on 26, 27)

A sterilisation method according to any one of claim 26 or 27 wherein the sterilisation method is a terminal sterilisation method.

### Claim 29 (depends on 26, 27, 28)

A sterilisation method according to claims 26 to 28 wherein the fluid is carbon dioxide.

### Claim 30 (depends on 26, 27, 28, 29)

A sterilisation method according to any one of claims 26 to 29 wherein the sterilant fluid comprises one or more additives selected from: a peroxide, carboxylic acid, acid anhydride, ester or alcohol, or is a commercial additive package.

### Claim 31 (depends on 26, 27, 28, 29, 30)

A sterilisation method according to any one of claims 26 to 30 wherein the additive is present in an amount of between about 0.001% to about 2.0% based on the total volume of the sterilant fluid.

### Claim 32 (depends on 26, 27, 28, 29, 30, 31)

A sterilisation method according to any one of claims 26 to 31 wherein the material in need of sterilisation is from a human donor.

### Claim 33 (depends on 32)

A sterilisation method according to claim 32 wherein the material in need of sterilisation is from one or more donors.

### Claim 34 (depends on 32, 33)

A sterilisation method according to any one of claim 32 or 33 wherein the material in need of sterilisation is from one or more live donors or from one or more cadaveric donors or is a mixture of live and cadaveric donors.

### Claim 35 (depends on 26, 27, 28, 29, 30, 31, 32, 33, 34)

A sterilisation method according to any one of claims 26 to 34 wherein the sterilisation chamber is 20 to 200 litres, 60 to 150 litres, or 80 to 100 litres.

### Claim 36 (depends on 26, 27, 28, 29, 30, 31, 32, 33, 34, 35)

A sterilisation method according to any one of claims 26 to 35, wherein the material in need of sterilisation has been pre-treated with a SCF, or wherein the material in need of sterilisation has been pre-treated with a SCF and wherein that SCF was SCCD.

### Claim 37 (depends on 36)

A sterilisation method according to claim 36 wherein the pre-treatment has removed 25 to 99% of the mass of the material pre-treated, or has removed 70 to 98% of the mass of the material pre-treated, or has removed 85 to 95% of the mass of the material pre-treated.

### Claim 38 (depends on 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37)

A sterilised material obtained by the sterilisation method of anyone of claims 26 to 37, or article derived thereof.

### Claim 39 (depends on 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38)

A sterilised material obtained by the sterilisation method according to any one of claims 26 to 38, or article derived thereof, wherein the sterilised material has been sterilised under conditions sufficient to achieve at least a 6-log or 12-log reduction, in a population of one or more CFUs.

### Claim 40 (depends on 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37)

A sterilised material obtainable by the sterilisation method according to anyone of claims 26 to 37, or article derived thereof, wherein the sterilised material has been sterilised under conditions sufficient to achieve at least a 6-log or 12-log reduction, in the population of one or more CFUs, and wherein this reduction is verifiable.

### Claim 41 (depends on 38, 39, 40)

A sterilised material according to anyone of claims 38 to 40, or article derived thereof, wherein the material is selected from therapeutic grade tissue suitable for transplant, bone, demineralised bone, demineralised bone matrix, a paste comprising demineralised bone or demineralised bone matrix, whole cortical bone pieces, tendon, Achilles tendon, cartilage, ligament, skin, connective or musculoskeletal tissue or biological tissue suitable for implantation; natural or synthetic polymer, biomedical polymer, biodegradable polymer, synthetic human tissue analogue; biologically active molecules, pharmaceutically active compounds, pharmaceutical carriers, or pharmaceutical delivery vehicles; metals, alloys, medical equipment, instruments, prosthetics or implants.

### Claim 42 (depends on 38, 39, 40, 41)

A sterilised material according to anyone of claims 38 to 41, or article derived thereof, wherein the material comprises Cortical Cancellous Crunch.

### Claim 43 (depends on 38, 39, 40, 41, 42)

A sterilised material according to anyone of claims 38 to 42, or article derived thereof, wherein the article derived thereof is selected from an allograft, implant, stent, catheter, endoscope, prosthesis, joint replacement, medical scaffolding, suture, medical instrument, surgical instrument, drug delivery device or pharmaceutically active implant or microparticles.

### Claim 44 (depends on 38, 39, 40, 41, 42, 43)

Use of a sterilised material, or article derived thereof, according to anyone of claims 38 to 43 in a method of medical treatment.

### Claim 45 (depends on 38, 39, 40, 41, 42, 43)

A method of medical treatment comprising the use of a sterilised material, or article derived thereof, according to anyone of claims 38 to 43.

### Claim 46 (depends on 38, 39, 40, 41, 42, 43)

Use of a sterilised material, or article derived thereof, according to anyone of claims 38 to 43 in the manufacture of a product for use in implantation or transplantation.

### Claim 47

A pre-treatment method, wherein a material in need of sterilisation is pre-treated with a SCF, or wherein the material in need of sterilisation is pre-treated with a SCF and wherein that SCF is SCCD.

### Claim 48 (depends on 47)

A pre-treated material obtained or obtainable by the pre-treatment method of claim 47.

### Claim 49 (depends on 48)

A pre-treated material according to claim 48, wherein the pre-treatment method has removed 25 to 99% of the mass of the material pre-treated, or has removed 70 to 98% of the mass of the material pre-treated, or has removed 85 to 95% of the mass of the material pre-treated.

### Claim 50 (depends on 48, 49)

A sterilisation method comprising bringing the pre-treated material of any one of claim 48 or 49 in need of sterilisation into contact with a sterilant fluid, wherein optionally the sterilant fluid comprises a fluid at or near the supercritical pressure and temperature for that fluid.