WO2013033720A1

WO2013033720A1 - Storage system for biological material

Info

Publication number: WO2013033720A1
Application number: PCT/US2012/053697
Authority: WO
Inventors: Eugene CALHAU
Original assignee: Bayer Healthcare, Llc
Priority date: 2011-09-02
Filing date: 2012-09-04
Publication date: 2013-03-07
Also published as: US20130056480A1

Abstract

An apparatus and kit for storing frozen biological material, the apparatus and kit comprising a storage container comprising at least one wall; a product container, the product container comprising a material compatible with biological material; and a pad. A method of storing biological material, the method comprising: inputting biological material into a product container, the product container comprising a bag, the bag comprising a material compatible with the biological material; freezing the biological material at a temperature in the range of about 0°C and -100°C; positioning the product container in a storage container; positioning at least one pad between the product container holding the frozen biological material and the storage container; storing the storage container, product container, and frozen biological material at a temperature sufficient to maintain the biological material in a frozen state.

Description

STORAGE SYSTEM FOR BIOLOGICAL MATERIAL

[001] This application is being filed on 4 September 2012, as a PCT

International Patent application in the name of Bayer HealthCare, LLC, a U.S. national corporation, applicant for the designation of all countries except the US, and Eugene Calhau, a citizen of the U.S., applicant for the designation of the US only, and claims priority to U.S. Patent Application Serial No. 13/225,026, filed September 2, 2011, the subject matter of which is incorporated by reference in its entirety.

Background

[002] Because of the numerous benefits offered by disposable or "single use" technology, it has become an integral part of researching and manufacturing biological materials such a biopharmaceuticals. Single use technology eliminates the need to clean, validate, and maintain multi-use storage systems. By using this technology, researchers and manufacturers of biological materials can lower cost, save processing time, and utilize processing flexibility. In addition, single-use technology often provides for efficient and space-saving storage of biological materials. For example, multi-use stainless steel cryo vessels require much more space to store than flexible plastic bags.

[003] Both researching and producing biological materials is expensive. To optimize use of research and production facilities, biological materials are produced in bulk and often stored at frozen temperatures for extended periods of time until market demand is such that the bulk biological material is thawed, further purified and formulated for use in research or in production of the final product for commercial sale. Because single-use technology has several advantages, many manufacturers prefer to freeze liquid biological materials in single-use containers. In some cases, these biological materials are stored and transported at temperatures as low as -100°C. Unfortunately, single-use containers are often plastic materials which become particularly fragile at lower temperatures. In addition, the freezing and thawing techniques used for the bulk biological materials also can adversely affect these types of container materials. [004] Damage to single-use containers also can occur during handling, particularly during transport and warehousing. In one study, single-use containers, in the form of plastic bioprocess bags, were subject to extensive manipulation to assess container resiliency under simulated processing conditions. Only bags that did not leak biological material after testing were considered to survive the test. The study determined that even the most resilient bags only survived 50% of the time during testing. Kilburn et al., "Evaluating Single-Use Frozen Storage Systems." Am. Pharmaceutical Review, April 12, 2010, pp. 12-18.

[005] Damage to these types of single-use containers often is not discovered until the product is thawed. As a result, manufacturers often have difficulty with, among other things, production scheduling and planning, and with unnecessary costs associated with storage of unusable product. Additionally, damage to the single-use container also means sterility of the biological material has been compromised making it unsuitable for use and resulting in the loss of a valuable material. Although some researchers and manufacturers have implemented process improvements and procedural controls to improve handling techniques, damage to single-use containers still occurs.

Summary

[006] An apparatus for storing biological material comprises a storage container. The storage container comprises at least one wall and defines a first void. A product container is removably positioned within the first void. The product container comprises at least one wall and defines a second void, which is at least partially filled with frozen biological material. At least a portion of a pad is positioned between at least a portion of the at least one wall of the storage container and at least a portion of the at least one wall of the product container.

[007] A method of storing biological material comprises inputting biological material into a product container, the product container comprising a bag, the bag comprising a material compatible with the biological material; freezing the biological material at a temperature in the range of about 0°C and to about -100°C; positioning the product container in a storage container; positioning at least one pad between the product container holding the frozen biological material and the storage container; and storing the storage container, product container, and frozen biological material at a temperature sufficient to maintain the biological material in a frozen state.

[008] A kit for storing frozen biological material comprises a storage container, a product container, and a pad. The storage container comprises at least one wall. The product container comprises a material compatible with biological material. The storage container, product container, and pad comprise only materials suitable for use at temperatures as low as about -100°C.

[009] These and other features are disclosed and taught herein.

Description of the Drawings

[010] The skilled artisan will understand that the drawings, described below, are for illustration purposes only. The drawings may not be to scale. The drawings are not intended to limit the scope of the present teachings or appended claims in any way.

[011] Figure 1 is and exploded isometric view of a product container and padding cut outs.

[012] Figure 2 is a cross sectional view of the product container illustrated in Figure 1, taken along line 2-2.

[013] Figure 3 is a cross sectional view of a pad illustrated in Figure 1, taken along line 3-3.

[014] Figure 4 is an isometric view of a side pad illustrated in Figure 1.

[015] Figures 5a-5d are isometric views of a storage container for holding the pads and product container illustrated in Figure 1.

[016] Figure 6 is a top plan view of an alternative storage container.

Detailed Description

[017] Various embodiments will be described in detail with reference to the drawings, wherein like reference numerals represent like parts and assemblies throughout the several views. Reference to various embodiments does not limit the scope of the claims attached hereto. Additionally, any examples set forth in this specification are not intended to be limiting and merely set forth some of the many possible embodiments for the appended claims.

[018] For purposes of this patent document, the terms "or" and "and" shall mean "and/or" unless stated otherwise or clearly intended otherwise by the context of their use. The term "a" shall mean "one or more" unless stated otherwise or where the use of "one or more" is clearly inappropriate. The terms "comprise," "comprises," "comprising," "include," "includes," and "including" are

interchangeable and not intended to be limiting. For example, the term "including" shall be interpreted to mean "including, but not limited to." Additionally, all ranges provided herein include the upper and lower values of the range unless explicitly noted.

[019] Referring now to Figure 1, a storage system 10 includes a product contamer 12 and padding 14 positioned to protect the product container 12. The product container 12 holds a biological material 16 that can be frozen at extremely low temperatures for storage. The product container 12 can become fragile and susceptible to damage when exposed to the very low temperatures commonly used to freeze the biological material 16. The padding 14 is a protective layer proximal to at least a portion of an outer surface of the product container 12. The padding 14 provides cushioning that protects the product container 12 by mitigating the effect of force and shock. The padding 14 also can provide thermal insulation to maintain the temperature of the product container 12 and the biological material 16, or at least prevent substantial warming of the product container 12 and biological material 16, for a period of time when it is not stored in a cold environment such as a freezer. Any one or combination of the product container 12 or padding 14 can be configured or designated for single use or for multiple use.

[020] Referring now to Figures 1 and 2, the product container 12 can be a bioprocess bag and can be filled with a biological material 16. The product container 12 and biological material 16 are typically frozen at very low temperatures when stored. The bioprocess bag can be formed with first and second opposing walls or panels 18 and 20 that define a void 26 for holding the biological material 16. The perimeter or outer perimeter of the first and second panels 18 and 20 forms four side portions 22a-22d. A first tube 24 defines a passage 25 and provides a port for inputting and extracting biological material 16 into and from the void 26 in the product container 12. The first tube 24 can be flexible and can include a valve or other mechanism to seal the passage 25. A second tube 28 also defines a passage 27 and provides a vent into the void 26 to enable easier flow of the biological material 16 though the first tube 24. The second tube 28 can be flexible and can include a valve or other mechanism to seal the passage 27. The product container 12 and the biological material 16 are discussed in more detail herein.

[021 ] When biological material 16 is filled into the void 26 of the product container 12, it causes the opposing panels 18 and 20 to separate, which can change the overall depth between the side panels 18 and 20 and the length of the side portions 22a-22d. The width, length, and depth of the product container 12 can vary depending on factors such as the volume of biological material 16 stored in the product container.

[022] Alternative embodiments of the product container 12 are possible and any suitable structure for inputting and extracting the biological material 16 can be used. For example, the first and second tubes 24 and 28 could be rigid. In another example, the product container 12 can define a single opening for inputting and extracting the biological material 16. The product container 12 can have any suitable size and dimensions. In at least one possible embodiment, the product container 12 is sized to hold about 5 liters of the biological material 16. At least some alternative embodiments are disclosed in United States patent application serial number 13/225,026, which was filed on September 2, 2011 and entitled Biopharmaceutical Product Storage System, the entire disclosure of which is hereby incorporated by reference.

[023] Each of the opposing panels 18 and 20 can have two layers of material such as an outer backing layer 30 and an inner contact layer 32 that comes into contact with the biological material 16. These layers 30 and 32 can be assembled using any method sanctioned by the biological material 16 manufacturer such as tie layer adhesives. The backing layer 30 can be configured to provide dimensional stability and structural support to the contact layer and the product container 12. It can have any thickness suitable to provide structural stability and maintain flexibility. The thickness also can vary depending on other factors such as how the product container 12 is intended to be handled and used and the environment in which the product container 12 is intended for use. The outer layer 30 is compatible with the biological material 16. [024] The contact layer 32 comes in contact with the biological material 16 and is compatible with the biological material 16 such that the biological material 16 does not unacceptably degrade, react, or absorb when in contact with the contact layer 32. The contact layer 32 has a thickness that prevents unacceptable effects on the biological material 16 over long periods. In at least one embodiment, the contact layer 32 has a thickness of about 360 μιη or more. However, the thickness of the contact layer 32 can vary depending on a variety of factors including the type of biological material that the product container 12 is designed to hold, the density of the biological material, and the environment in which the product container 12 will be stored. Other embodiments of the contact layer 32 can have other thicknesses. Other structural requirements for the contact layer 32 may include density and tensile strength. For example, one or more embodiments of the contact layer 32 may have a density of about 0.94 g/cm3 or more. In another example, at least one embodiment of the contact layer 32 may have a tensile strength at about 100% elongation of about 6 MPa (870 psi) or more and an elastic modulus of about 50

MPa (7.25 kpsi) or more. At least some of these properties may be measured, using accepted industry standards, including ISO 527-2. Tests that can be used to determine whether the backing layer 30 and the contact layer 32 are compatible with a particular biological material include tests performed according to U.S.

Pharmacopeia Reference Standards.

[025] Although two layers 30 and 32 are disclosed, alternative

embodiments can use a single layer of material or more than two layers of material to form the panels 18 and 20 of the product container 12.

[026] To verify and validate that the product container 12 is compatible with the biological material 16 and meets other design requirements, the material used to form the layers 30 and 32 and the product container 12 itself can be tested for product stability, product adsorption, gas permeability, sterilization, chemical compatibility, leachables, extractables, and the like. At least some of the testing can be performed by inserting biological material into product containers 12 and then measuring effects of the product containers 12 on the biological material 16 over a specified duration. The material used to form the layers 30 and 32 and the product container 12 may be tested for strength and product integrity. The parameters that the material forming the layers 30 and 32 and other components of the product container 12 should meet may be varied depending on manufactures specifications, customer specifications, the biological material 16 to be stored in the product container 12, the environment in which the product container 12 is to be used and stored, and the like.

[027] Examples of materials that can be used to form the backing layer 30 and the contact layer 32 include ethylene and vinyl acetate (EVA); ethyl vinyl alcohol (EVOH); polyvinylidene fluoride (PVDF); polyethylene and polypropylene (low density and high density); polytetrafluoroethylene; silicone; nylon; polyesters; polyolefin homopolymers; other polyolefins; thermoplastics; and polymeric materials; combinations thereof; and the like. The various layers 30 and 32 of the product container 12 can be formed with other natural on synthetic compounds, that are not reactive, additive, or absorptive such that the purity, strength, or identity of the biological material 16 held in the product container 12 is compromised.

[028] Examples of product containers 12 include those manufactured by

Arkema, Inc., DuPont, Dow Chemical Company, Sartorius-Stedim Biotech S.A., and Charter Medical, Ltd., among other manufacturers.

[029] Referring back to Figure 1, the padding 14 can include six panels including a lower pad 34, an upper pad 36, and four side pads 38a-38d. In use, the lower pad 34 is positioned proximal to one panel 18 or 20 of the product container 12 and the upper pad 36 is position proximal to the opposing panel 20 or 18, respectively, of the product container 12. In at least one possible embodiment, the lower and upper pads 34 and 36 have about the same dimensions as the first and second panels 18 and 20 of the product container 12. Each of the four side pads 38a-38d is positioned proximal to one of the side portions 22a-22d, respectively, the product container 12. In at least one embodiment, each side pad 38a-38d has a length that is about the same length as the side portion 22a-22d, respectively, of the storage container 12 to which it is proximally positioned.

[030] Referring now to Figures 1 and 3, each pad 34, 36, 38a-38d may be wrapped or enclosed in a protective cover or layer 54, 56, 58, respectively. For example, the upper pad 36 may be wrapped in a protective cover 56 formed with a single sheet of material 42. The sheet of material 42 is folded over the upper pad 36 so a first half of the sheet 42 covers a top surface 44 of the upper pad 36 and a second half of the sheet 42 covers a bottom surface 46 of the upper pad 36. The opposing peripheral or edge portions 48a-48c of the first and second halves of the sheet 42 can then attached to one another to enclose or seal the pad 36 within the protective cover 54. The opposing peripheral edge portions 48a-48c of the sheet of material 42 forming the protective covering 54 can be attached to one another by any suitable means such as an adhesive, heat sealing, or any other mechanical structure or mechanism suitable for the material of the protective covering 54 and the environment in which the upper pad 36 is to be used. The lower pad 34 can be similarly sealed in the protective cover 56.

[031] Referring to Figures 1 and 4, all four side pads 38a-38d can be similarly wrapped in the protective cover 58, which also can be formed with a single sheet of material. To cover the side pads 36a-36d, they are positioned end-to-end on the single sheet with a slight gap 50a-50c between the ends of adjacent side pads 36a-36d. The single sheet of material is then folded over the top surface of the side pads 36a-36d so that a first half of the sheet covers the bottom surface of the side pads 36a-36d and a second half of the sheet covers the opposite or top surface of the side pads 36a-36d. The opposing peripheral edges of the first and second halves of the sheet are then attached to one another, respectively, to enclose the side pads 36a- 36d to form the protective cover 58. Additionally, the portions of the two opposing halves of the sheet that extend along the gaps 50a-50c between adjacent side pads 36a-36d can be attached to each other to form an intermediate section 52a-52c between adjacent side pads 36a-36d. Each intermediate section 52a-52c maintains the gap 50a-50c, respectively, between adjacent side pads 36a-36d and forms a living hinge so adjacent side pads 36a-36d can rotate around the intermediate section 52a-52c and one another.

[032] In at least one possible embodiment, the material used to form the protective covers 54, 56, 58 for the pads 34, 36, 38a-38d, respectively, is a heat shrink material. In these embodiments, the protective covers 54, 56, 58 can be heated after the pads 34, 36, 38a-38d are enclosed. Heating the protective covers 54, 56, 58 will cause the material to shrink around the pads 34, 36, 38a-38d to minimize any waviness or folds in the protective covering 54, 56, and 58. [033] The material forming the protective covers 54, 56, 58 is compatible with the material used to form the product container 12 and is suitable for direct contact during handling by people and apparatuses that may come into contact with it. The protective covers 54, 56, 58 can be made from a resilient material that is resistant to tears, cracks, and punctures. In at least one possible embodiment, the material forming the protective covers 54, 56, 58 for the pads 34, 36, 38a-38d, respectively, has a shear strength in the range from about 124 MPa (18 kpsi) to about 150 MPa (22 kpsi) and maintains dimensional stability over a wide temperature range from about -70°C to about 150°C and, under certain conditions, is suitable for use at temperatures from about -250 °C to about 200°C. An example of material that can be used to form the protective covers 54, 56, 58 includes polyester based material such as biaxially-oriented polyethylene terphthalate ("BoPet"). One commercially available type of BoPet is Mylar® brand polyester film, which is manufactured by DuPont Teijin Films. Other materials that can be used to form the protective covers 54, 56, 58 include plastics, biodegradable materials, and other materials other than polyester films.

[034] An advantage the protective covers 54, 56, 58 is that they protect the product container 12 from damage if any of the pads 34, 36, 38a-38d are formed with a material that may be abrasive or might otherwise damage the product container 12 if the pad and product container 12 come into contact with each other. In at least one alternative embodiment, one or more of the pads 34, 36, and 38a-38d do not include or are not otherwise wrapped in any protective covers.

[035] Referring now to Figures 1 and 3, the pads 34, 36, 38a-38d can be formed with one or more mineral fibers 60 made from rock and/or slag. For example, the rock material can be a basalt rock. Basalt rock is a volcanic rock generally comprising plagioclase, pyroxene, and olivine. Before formation into a mineral fiber, basalt rock is hard and dense with a glassy appearance. After the rock is formed into a fiber, its density ranges from about 2.7 g/cm³ to about 2.9 g/cm³. Slag materials are byproducts of various types of metallurgical operations. These materials are generally non-metallic and comprise oxides of silica, lime, alumina and magnesia. After the slag is cooled and solidified, it may be spun to form mineral fibers. Where the fibers are manufactured from a combination or basalt rock and recycled slag, the rock and slag are heated to approximately 1540°C (2000°F), until in a molten state. After the molt is cooled and solidified, it is spun to form mineral fibers. The fibers are used to form each of the pads. An example of mineral fiber pads enclosed in a protective covering includes CONTROL TEMP PACKAGING®- brand material, which is commercially available from R.N.C. Industries Inc., Norcross, GA.

[036] In alternative embodiments, the mineral fibers 60 can be formed with material other than Basalt and/or slag. The pads 34, 36, 38a-38d can be formed with protective material other than mineral fibers 60 that protects the product container 12 against damage and/or provides a thermal insulator. Polystyrene foam is an example of such a material. Additionally, the various pads 34, 36, 38a-38d that form the padding 14 can be formed with different material such that one pad is formed with one material and another pad is formed with a different material. The pads 34, 36, 38a-38d can be formed with materials that are eco-friendly,

biodegradable, generally semi-rigid, rigid, and/or flexible, and the like.

[037] Different factors can affect the type of material used to form the pads

34, 36, 38a-38d and the thickness of the pads 34, 36, 38a-38d, including the environment in which the pads 34, 36, 38a-38d and storage system 10 are to be used, the amount of handling expected for the pads 34, 36, 38a-38d and the storage system 10, the amount of cushioning or protection desired to protect the product container 12 from damage, and the temperature specifications required to maintain the biological material 16 in a frozen state when the storage system 10 is not refrigerated. In at least one possible embodiment, the pads 34, 36, 38a-38d have a thickness of about 1 inch (2.54 cm), about 1.5 inches (3.81 cm), about 2 inches (5.08 cm), or greater. The pads 34, 36, 38a-38d also can have a thickness less than about 1 inch (2.54 cm). The top, bottom, and side pads 34, 36, 38a-38d can have the same thickness or different thicknesses.

[038] Referring now to Figures 5 A and 5D, a storage container 62 can be box-like and has a bottom panel 64. First and second side walls or panels 66a and 66b can be positioned at opposite side edges of the bottom panel 64. The first and second side walls 66a and 66b can each have a leading edge portion 68a and 68b, respectively, orientated orthogonally to the bottom panel 64 and facing the front portion of the storage container 62. Slots 70a and 70b can be defined in the leading edge portions 68a and 68b, respectively, of the first and second side walls 66a and 66b, respectively. A front wall or side panel 72 is positioned at the leading edge of the bottom panel 64 and can extend between the first and second side walls 66a and 66b. A rear wall or panel 74 can be positioned at the read edge of the bottom panel 64 and extends between the first and second walls 66a and 66b. The bottom panel 64, the first and second side walls 66a and 66b, the front wall 72, and the rear wall 74 define a void 76.

[039] A top panel 78 can form a lid 80 for enclosing the void 76. The top panel 78 has a rear edge portion 81 operably connected to the rear wall 74 to form a living hinge so that the lid 80 can rotate to selectively open and close the storage container 62. A front flap 82 is operably connected to a front edge portion of the top panel 78 to form a living hinge so the front flap 82 can rotate relative to the top panel 78 and be positioned against the front wall 72 of the storage container 62 when the lid 80 is closed.

[040] First and second side flaps 83a and 83b can be rotatably connected to opposite side edges of the top panel 78 to form living hinges. The first and second side flaps 83a and 83b can be inserted between the first and second side walls 66a and 66b of the storage container 12, respectively, and the first and second side pads 38a and 38b, respectively. First and second closure flaps or tabs 84a and 84b can be rotatable connected to opposite side edges of the front flap 82 to form a living hinges. The first and second closure flaps 84a and 84b are positioned so they can be inserted into the slots 70a and 70b, respectively, defined in the first and second side walls 66a and 66b, respectively, to hold the lid 80 in a closed state. Although side flaps are disclosed for holding the lid 80 in a closed state, alternative embodiment could use adhesives, string tie downs, hook and loop fasteners such as VELCRO® brand fasteners, or any other suitable fastening structure, or combinations thereof.

[041] In at least one possible embodiment, the storage container 62 can be formed with a corrugated plastic such that each panel and flap forming the storage container 62 comprises flutes sandwiched between two or more layers of plastic sheets. An advantage of using corrugated material to form the storage container 62 is that it provides additional cushioning to supplement the cushioning provided by the padding 14, but is still rigid enough to substantially maintain its shape.

Alternative embodiments of the storage container 62 can be made from other materials such as cardboard, wood, metal, stone, Styrofoam, mineral fiber similar to the pads, corrugated materials, non-corrugated material, and the like.

[042] Although the storage container 62 is illustrated having a rectangular box-like shape, other shapes and configurations are possible. For example, the storage container 62 could be round, oblong, triangular, hexagonal, or any other suitable shape. In another example, the storage container could be formed from a material that has a low enough structural rigidity to form a bag, wrap, or similar structure suitable to hold a pad in place relative to the product container 12. The storage container 62 can be configured or designated for single use or for multiple use.

[043] Referring to Figure 6, a bag 86 can form the storage container in at least one alternative embodiment and can have first and second opposing panels 88 and 90. Three outer edge portions 92a-92c along the periphery of the first panel 88 are attached to the three opposing outer edge portions 92a-92c of the second panel 90. The fourth opposing outer edge portions 92d of the panels 88 and 90 define an opening 94 into a void 96 formed between the first and second panels 88 and 90. A flap 98 can be positioned along the fourth outer edge of either the first or second panel 88 or 90 and can be folded over and secured to the other of the second or first panels 90 or 88, respectively. The securing mechanism can be any known mechanism such as an adhesive, tabs, or string tie-downs. Hook and loop fasteners, or any other suitable mechanism. The bag 86 can be made of any suitable material such as paper, plastic, cloth, polyester film, any other suitable material, and combinations thereof.

[044] The storage container 62 can have any dimensions suitable for holding the product container 12 and one or more pads positioned to protect the product container 12. In one example, the storage container 62 has dimensions about 20 inches (50.8 cm) by about 16 inches (40.64 cm) by about 3 inches (7.62 cm). In another example, the storage container 62 has dimensions of about 21 inches (53.54 cm) by about 59 inches (149.86 cm). [045] In at least one embodiment of the storage system 10 does not have a separate storage container 62 and padding 14. In these embodiments, the storage container can be formed with the pads themselves. For example, a bottom panel, a top panel, and side walls of a box-like storage container can be formed with pads including the mineral fiber pads and polystyrene foam pads described herein. In another example, a storage container is formed with pads having a low enough structural rigidity such that the pads can form a bag, wrap, or other suitable structure to hold the product container 12 and provide cushioning to mitigate the effect of force and shock against the storage container has on the storage container 12.

[046] A label (not shown) containing information can be applied to an outer surface of the storage contain 62 to convey information without having to open the storage container 62. The label can contain information to identify the biological material 16 being stored in the product container 12, the volume of the biological material 16, storage temperatures, dates, warnings, and any other information.

[047] In use, the four side pads 38a-38d can be inserted into the storage container 62 and positioned so that they rest on the top surface of the bottom panel 64 for the storage container 62. The first and second side pads 38a and 38b can be positioned adjacent to the first and second side walls 66a and 66b, respectively, of the storage container 62 and have substantially the same height and length as the first and second side walls 66a and 66b, respectively. The front and rear side pads 38c and 38d also can be positioned adjacent to the front and rear side walls 72 and 74, respectively, of the storage container 62 and have substantially the same height and length as the front and rear side walls 72 and 74, respectively. At least two opposing side pads (e.g., first 38a and second 38b, or front 38c and rear 38d) can be slightly shorter than their adjacent side wall 66a, 66b, 72, and 74, respectively, to make room for the end portions of the two other opposing side pads (e.g., front 38c and read 38d, or first 38a and second 38b).

[048] The lower pad 34 can be positioned against the bottom panel 64 of the storage container 62. In this position, the four side pads 38a-38d are adjacent to and extend around the outer edge or periphery or circumference of the lower pad 34.

[049] As illustrated in Figure 5B, the product container 12, which is at least partially filled with frozen biological material 16, then can be positioned in the void 76 of the storage container 62 and positioned so that is rests on the lower pad 34. The product container 12 and side pads 38a-38d can be sized so each outer edge portion 22a-22d of the product container 12 is positioned proximal to one of the four side pads 38a-38d to minimize movement of the product container 12 relative to the lower pad 34. The side pads 38a-38d and the product container 12 can be sized so the side edges 22a-22d of the product container 12 do not press against the side pads 38a-38d with enough force that the product container 12 is damaged. How snuggly the side portion of the product container 12 meet against the side pads, or whether the side portions of the product container 12 even rests against their adjacent side pads, can vary depending on a variety of factors such as the volume of biological material 16 held in the product container 12.

[050] The first and second tubes 24 and 28 can be folded so that the entire length of the top edge 22c of the product container 12 is positioned against, or at least proximal to, its adjacent side pad 38c. Various embodiments might include a hole or recess in the side pad 38c that is sized and positioned to receive the first and second tubes 24 and 28 so that the entire top edge 22c of the product container 12 can be positioned against, or at least proximal to, the side pad 38c without bending the tubes 24 and 28.

[051 ] Referring now to Figure 5C, the upper pad 36 can be positioned over the product container 12 and positioned so the four side pads 38a-38d are adjacent to and extend around the outer edge or periphery or circumference of the upper pad 36. In at least one embodiment, the combined thickness or depth of the lower pad 34, upper pad 36, and product container 12 can be about the same height as the side walls 66a, 66b, 68a, and 68b of the product container 12 which may minimize movement of the product container 12 up and down or relative to the height of the side walls 66a, 66b, 68a, and 68b of storage container 62. The combined thickness of the lower pad 34, upper pad 36, and product container 12 may not be so thick that the lid 80 of the storage container 62 exerts enough force against the upper pad 36 to damage the product container 12. The combined thickness of the lower pad 34, upper pad 36, and product container 12 can vary depending on factors such as the volume of biological material 16 held in the product container 12. [052] Alternative embodiments might use padding formed with more or less than six pads. For example, a single pad can be positioned on the bottom panel 64 of the storage container 62 and the product container 12 can then rest on the single pad. In another example, a pad can be positioned on the bottom panel 64 of the storage container 62 and one or more side pads positioned along the side walls 66a, 66b, 72, and 74 of the storage container, while no top pad is used. In yet another example, only top and bottom pads are used.

[053] Alternative embodiments might use different shapes, configurations, and arrangements for the pads. For example, each pad can be sized smaller (e.g., width and/or length) than the panels 64 or 78 or side walls 66a, 66b, 72, and 74 of the storage container 62 against which they are positioned. In other examples, two or more pads might be placed side-by-side along the surface of the bottom panel 64 of the storage container 62 or two or more pads and can be layered to increase the thickness of the padding. Another example is that one or more of the pads may have shapes other than rectangles; may have a contour so they are flat or planer so their entire surface does not lay against their adjacent panel or wall of the storage container 62 or product container 12; or may define holes, recesses, or voids such that they do not cover the entire surface of a panel or wall of the storage container 62. In another example, a pad has one surface that is flat for positioning against a panel or wall of the storage container 62 and an opposite surface that is contoured to the shape of the product container 12.

[054] If a bag-like storage container 86 is used, such as the storage container illustrated in Figure 6, the product container 12 can be inserted into the bag-like storage container 86 together with at least one pad to protect the product container 12. The product container 12 can be protected my more than one pad as appropriate. For example, the product container 12 can be protected by two pads— one pad positioned adjacent to one panel 18 of the product container 12 and another pad positioned adjacent the opposing panel 20 of the product container 12. In another example, the product container 12 can be protected by more than two pads such that a pad is positioned adjacent to one panel 18 of the product container 12, a pad is positioned adjacent the opposing panel 20 of the product container 12, and a pad is positioned adjacent each of the side edges 22a-22d of the product container 12.

[055] As used herein, biological material 16 refers to any biological material, biopharmaceutical product, intermediaries thereof, and derivatives thereof that changes, at some point during processing, from a liquid to a frozen state. The temperature of the biological material while in the liquid state typically is maintained from about 2°C to about 8°C. The temperature of the biological material while in the frozen state typically ranges from about -100°C to about 0°C.

Specifically, the biological material may be handled, stored, and transported at temperatures at or slightly below about 0, about -10, about -20, about -30, about -40, about -50, about -60, about -70, about -80, about -90 or about -100°C or at a temperature falling within any combination of these temperatures as an upper and lower limit, such as at about 0 to about -10°C. Temperatures of the biological material 16 while in either the liquid or frozen state may also be lower or higher than those stated, depending on the properties and processing specifications of the biological material 16.

[056] The biological material 16 may comprise a liquid solution of recombinant proteins, antibodies (monoclonal or otherwise), vaccines,

blood/plasma-derived products, nonrecombinant culture-derived proteins, and cultured cells. The liquid solution of recombinant protein may comprise a solution of any recombinant protein obtained from recombinant cell culture and isolated at least partially from the cell culture medium using affinity chromatography, ion- exchange chromatography, or the like. As used herein, "solution" includes suspensions, dispersions and the like of the biological material in a liquid vehicle.

[057] The solution may comprise a bulk solution, which is a solution which has been partially purified. As used herein, a "bulk" solution comprises a partially but not fully purified liquid solution of biological material such as a recombinant protein. Bulk solutions are further characterized by their very low product concentration. In some embodiments, the solution may be of a biological material such as a recombinant protein at about 0.0001 micromolar, 0.001 micromolar, about 0.01 micromolar, or a range between about 0.0001 to about 0.001 micromolar or about 0.001 to about 0.01 micromolar. In some embodiments of the invention, the concentration of biological material such as a recombinant protein can be as high as about 10 micromolar, about 1.0 micromolar, or about 0.1 micromolar, or a range between about 10 to about 1.0 micromolar or about 1.0 to about 0.1 micromolar or about 0.1 to about 0.01 micromolar or about 10 to about 0.01 micromolar or 10 to 0.0001 micromolar or 10 to 0.001 micromolar or 10 to 0.1 micromolar or any other concentration falling between any combination of these upper and lower limits of protein concentration.

[058] Often during production of recombinant proteins, an elution buffer of high salt content is used to elute the desired protein from a first-pass purification treatment. In the case of elution from a column, a high salt concentration is needed to release the protein from the column. Accordingly, the bulk solution recovered from first pass purification treatment can comprise a solution having a high concentration of monovalent salts, normally sodium chloride but potentially potassium chloride or other salts. The concentration of sodium chloride or potassium chloride in some embodiments is at least about 100 millimolar, at least about 200 millimolar, at least about 300 millimolar, at least about 400 millimolar, at least about 500 millimolar, at least about 600 millimolar, at least about 700 millimolar, or at least about 800 millimolar. The bulk solution may also contain varying amounts of other salts, such as divalent salts, including calcium chloride. By "partially but not fully purified" is meant the liquid solution has been subjected to at least one purification step, but the liquid solution still contains sufficient residual impurities that at least one further purification step is required prior to final product formulation. For example, a "bulk" solution of recombinant Factor VIII must be further purified prior to final formulation, which in the case of Factor VIII and other proteins may include lyophilization.

[059] Recombinant proteins include, for example and without limitation, coagulation factors, virus antigens, bacterial antigens, fungal antigens, protozoal antigens, peptide hormones, chemokines, cytokines, growth factors, enzymes, blood proteins such as hemoglobin, a- 1 -antitrypsin, fibrinogen, human serum albumin, prothrombin/thrombin, antibodies, blood coagulation and/or clotting factors, and biologically active fragments thereof, such as Factor V, Factor VI, Factor VII, Factor VIII and derivatives thereof such as B-domain deleted FVIII, Factor IX, Factor X, Factor XI, Factor XII, Factor XIII, Fletcher Factor, Fitzgerald Factor, and von Willebrand Factor; milk proteins such as casein, lactoferrin, lysozyme, -l- antitrypsin, protein factors, immune proteins, and biologically active fragments thereof; and antibodies, including monoclonal antibodies, single chain antibodies, antibody fragments, chimeric antibodies, humanized antibodies, and other antibody variant molecules which can be produced in recombinant cell culture.

[060] The biological material may be derived from cell culture. The cell culture may comprise any type of cell including a plant, insect, mammalian, yeast or bacterial cell. In one embodiment, the biological material is an

ultrafiltered/diafiltered (UF/DF) solution obtained from cell culture. In another embodiment the biological material is an ultrafiltered/tissue culture concentrated filtered (UF/TCF) solution obtained from cell culture.

[061] In one embodiment, the biological material is recombinant Factor

VIII. Factor VIII as used herein includes engineered variants of Factor VIII, such as B-domain deleted variants of Factor VIII and site-specific mutation variants of Factor VIII or of B-domain deleted Factor VIII. The biological material may be a derivative of Factor VIII having Factor VIII procoagulant activity.

[062] In use, biological material 16 while in the liquid state is typically maintained in a temperature range from about 2°C to about 8°C. Biological material 16 while in the frozen state is typically maintained in a temperature range from about 0°C to about -100°C. In various embodiments, the biological material 16 when in the frozen state can be handled, stored, and transported at temperatures at or slightly below about 0, about -10, about -20, about -30, about -40, about -50, about - 60, about -70, about -80, about -90 or about -100°C or at a temperature falling within any combination of these temperatures as an upper and lower limit, such as at about 0 to about -10°C. Temperatures of the biopharmaceutical product while in either the liquid or frozen state also may be lower or higher than those stated, depending on the properties and processing specifications of the biological material 16.

[063] When processed for storage, the biological material 16 is input or filled into the product container 12 and the product container 12. The ports in the product container 12, (e.g., first and second tubes) are then sealed. The biological material 16 then can be frozen depending on the type of material it is and the timing of its intended use.

[064] To freeze the biological material 16, the product container 12 holding the biological material 16 can be placed in blast freezer to quickly lower the temperature of the biological material 16 to a desired or predetermined temperature. The blast freezer can use any suitable technique to freeze the biological material 16. For example, the biological material 16 can be quickly frozen by submersing the product container 12 in liquid nitrogen, a mixture of liquid nitrogen and water, a mixture of dry ice and ethanol, or any other material suitable for rapidly freezing the biological material 16 while it is in the product container 12. In various

embodiments, the blast freezer is capable of reducing the temperature of the biological material 16 and the product container 12 to a temperature as low as about -100°C. In at least one embodiment, the blast freezer maintains a temperature of about -56°C.

[065] After the temperature of the frozen biological material 16 is lowered to at least the desired temperature, the product container 12 can be removed from the blast freezer. The product container 12 is placed in the storage container 62 and protected by the padding 14. The storage container 62 can be transported to and placed in a long-term storage freezer so the biological material 16 can be maintained at a desired temperature for extended period of time. Storage at this temperature may be maintained for a long period of time such as periods up to about 30, about 60, about 90, about 180 days or longer. Often freezing occurs at one site in the manufacturing facility and the long term storage is at a second site some distance away.

[066] Transporting the product container 12 and frozen biological material

16 in the storage container 62 has several advantages. For example, the product container 12 holding the frozen biological material 16 also is at a very low temperature, usually the same temperature as the biological material 16, which may cause it to become fragile and susceptible to damage. In another example, at least some biological materials 16 are required to be maintained at a relatively stable temperature. For example, some biological material 16 may be required to be maintained at a relatively stable temperature of about -30°C or lower. The padding 14 can provide thermal insulation to maintain the temperature of the biological material 16, or at least prevent substantial warming of the product container 12 and biological material 16, for a period of time during transport or when not stored in a cold environment such as a freezer.

[067] The storage system 10 substantially prevents damage to the product container 12 that would result in exposure of the biological material 16 to adverse elements during product handling, storage, and transport. If the product container 12 is damaged, the storage system 10 also can prevent or minimize leakage of the biological material 16 when it is thawed while still in the product container 12. Protection of the product container 12 can be measured when, for example, at least 100 product containers 12 holding biological material 16 are subjected to a freeze- storage-thaw cycle and fewer than 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1% of the product containers 12 leak or are otherwise damaged during the freeze-storage-thaw cycle. This measurement may be made under any standard conditions of freeze-storage- thaw cycles known to one of skill in the art for storing the biological material 16.

[068] The biological material 16 can be thawed when ready for use, including further purification and formulation. As noted herein, the freeze-storage- thaw cycle can cause the temperature of the biological material 16 to vary in the temperature range from about -100°C to about 8°C. In other situations, the freeze- storage-thaw or the storage-thaw cycle is in the temperature range from about -30°C to about 2°C. Any suitable technique can be used to warm the product from a frozen state to a liquid state so it can be used. In at least one possible process, the biological material 16 can be thawed while the product container 12 is in the storage container 62. In at least another possible embodiment, the biological material 16 is removed from the storage container 16 and padding 14 before while it is being thawed. Agitation may be used to speed the thawing process although agitation can subject the product container 12 to damage. Accordingly, at least some processes do not use agitation while thawing the biological material 16 or use it only after the product container 12 reaches a temperature warm enough that it is not as susceptible to damage.

[069] All components and materials forming or related to the product container 12, padding 14, and storage container 62 are suitable for use in the temperatures for which they will be exposed such that the components and materials can be used for their intended purpose at room temperatures and at very low temperatures. For example, any adhesive used in the product container 12 is suitable for temperatures as low as those found in the blast freezer such that a seal is maintained. Examples of temperatures for blast freezing are disclosed herein and include -100°C. Adhesives and materials used in the storage container can be suitable for temperatures used in the blast freezing process as well. If components, such as the storage container 62 and padding 14, will be exposed to the long-term storage freezer and not the blast freezer, then then adhesives and other materials used in those components should be suitable for use at temperatures used for long- term storage. Examples of temperatures for long-term storage are disclosed herein.

[070] Each embodiment described herein is provided only for illustration and should not be used to limit the scope of the following claims. Other embodiments are possible for each of the structures, combinations of structures, actions, and combinations of actions recited in the following claims. Those skilled in the art will readily recognize various modifications and changes that may be made without following the example embodiments and applications illustrated and described herein, and without departing from the true spirit and scope of the following claims.

Claims

The claims are:

1. An apparatus for storing biological material, the apparatus comprising:

a storage container comprising at least one wall, the container defining a first void;

a product container removable positioned within the first void, the product container comprising at least one wall, the product container defining a second void, the second void being at least partially filled with frozen biological material; and

a pad, at least a portion of the pad positioned between at least a portion of the at least one wall of the storage container and at least a portion of the at least one wall of the product container.

2. The apparatus of claim 1 wherein the product container and the biological material is frozen at a temperature of about -30°C or lower.

3. The apparatus of claim 2 wherein the pad comprises mineral fiber.

4. The apparatus of claim 3 wherein the pad comprises mineral fiber having a density in the range of about 2.7 g/cm and about 2.9 g/cm .

5. The apparatus of claim 4 wherein the mineral fibers comprise basalt rock, plagioclase, pyroxene, olivine, slag, or combination thereof.

6. The apparatus of claim 4 wherein the pad is at least partially covered with a material, the material comprising a polymeric material, a biaxially-oriented polyethylene terephthalate, or combinations thereof.

7. The apparatus of claim 6 wherein the material has shear strength in the range of about 18 kpsi and about 22 kpsi.

8. The apparatus of claim 3 wherein the storage container comprises a bottom having an outer periphery, four side walls positioned around the outer periphery of the bottom, and a lid arranged to selectively open and close the first void.

9. The apparatus of claim 8 further comprising a plurality of pads including at least one pad between the bottom of the storage container and the product container, at least one pad between each of the side walls and the product container, and at least one pad between the lid and the product container.

10. The apparatus of claim 3 wherein the storage container comprises a bag.

11. The apparatus of claim 10 wherein: the product container comprises a bag; and

the bag comprises a material, the material being compatible with the frozen biological material.

12. The apparatus of claim 1 wherein the at least one wall of the storage container comprises the pad.

13. An apparatus for storing biopharmaceutical product, the apparatus comprising: a storage container defining a first void, the storage container comprising a bottom having an outer periphery, four side walls positioned around the outer periphery of the bottom, and a lid arranged to selectively open and close the first void;

a product container positioned in the first void, the product container

defining a second void, the product container comprising a bag at least partial formed with a material compatible with a biological material;

a plurality of pads positioned between the product container and the storage container, including at least one pad between the bottom of the storage container and the product container, at least one pad between each of the side walls and the product container, and at least one pad between the lid and the product container, one or more of the plurality of pads comprising micro fiber; and

the second void being at least partially filled with the biological material, the biological material being frozen at a temperature of about -30°C or lower.

14. A method of storing biological material, the method comprising:

inputting biological material into a product container, the product container comprising a bag, the bag comprising a material compatible with the biological material;

freezing the biological material at a temperature in the range of about 0°C and about -100°C;

positioning the product contamer in a storage container;

positioning at least one pad between the product container holding the frozen biological material and the storage container; and storing the storage container, product container, and frozen biological material at a temperature sufficient to maintain the biological material in a frozen state.

15. The method of claim 13 wherein the biological material is frozen in one freezer and the storage container, product container, and frozen biological material are stored in a second freezer.

16. The method of claim 13 wherein the first freezer is a blast freezer and the biological material is frozen by submersing the product container 12 in liquid nitrogen, a mixture of liquid nitrogen and water, a mixture of dry ice and ethanol, or any combination thereof.

17. The method of claim 14 wherein the product container holding the frozen biological material is transported to the second freezer in the storage container.

18. The method of claim 17 wherein the product container holding the frozen biological material is transported to the second freezer at a remote location from the first freezer.

19. The method of claim 13 further comprising removing the storage container from the second freezer and thawing the biological material.

20. The method of claim 19 wherein the product container is removed from the storage container before thawing the biological material.

21. A kit for storing frozen biological material, the kit comprising:

a storage container comprising at least one wall;

a product container, the product container comprising a material compatible with biological material; and

a pad;

wherein the storage container, product container, and pad comprise only materials suitable for use at temperatures as low as about -100°C.