WO2018004328A1

WO2018004328A1 - Device for the thermal protection and transport of biomacromolecules

Info

Publication number: WO2018004328A1
Application number: PCT/MX2017/000069
Authority: WO
Inventors: Abel MORENO CÁRCAMO; Claudia Carina PAREJA RIVERA
Original assignee: Universidad Nacional Autónoma de México
Priority date: 2016-06-29
Filing date: 2017-06-28
Publication date: 2018-01-04
Also published as: MX2016008614A

Abstract

Dewar flasks are containers used to transport materials that have to be kept at a low temperature and protected against vibration, shaking, impact, oscillation and/or any other external disturbance that could interfere with thermodynamic stability. However, if the Dewar flask is opened arbitrarily, such as for customs inspections during air transportation, without knowledge of the material contained therein, this could affect not only the low temperature inside the container, but also the intrinsic properties of the material therein. The device or container of the present invention is suitable for materials that have to be isolated from changes in temperature and/or external disturbances and can be used to transport said materials under suitable conditions from one site to another, including over long distances and intercontinental journeys, without any risk of damage from handling or external disturbances and without any thermal risk.

Description

The present invention relates to container devices for the protection and transport of materials, such as biological macromolecules, without risks of tampering or external disturbances, and without risks of thermal damage, from one site to another, even in transcontinental paths.

AND! The development of this invention was possible thanks to the sponsorship of the following projects: DGAPA UNA, IT2G0215 project and support from REDTÜLS (CONACYT Synchrotron Radiation Thematic Network of Users).

BACKGROUND

Gíbbs (1839-1903) was interested in the first and second principles of thermodynamics, focusing particularly on S entropy and its maximization a! evolve a closed system spontaneously. While it started with e! analysis of the thermodynamics of fluids, soon addressed the case in which different states of matter are present. Gibbs' contributions to the definition of phases, phase rule and equilibrium were crucial in chemistry, physics, metallurgy and materials, particularly for the systematization of the presentation and the analysis of the phase diagrams to equilibrium, concentration determination of crystal defects to balance, etc. A system is in thermodynamic equilibrium when it is in mechanical, thermal and chemical equilibrium. In materials it is particularly relevant e! aspect of chemical equilibrium. Jan! Within the Gibbs developments, it will be considered a solid, free to expand or contract, that has a given composition, and is at constant temperature T and pressure P. We will also impose that the material is not subjected to any external field. When analyzing the chemical equilibrium, not only the effect of the bonds must be taken into account, but also the entropic tendency of the atoms to mix (not only with each other, but also with the crystalline defects). Each previous effect can be expressed as an energy: the internal energy E and the mixing energy TS, respectively. On balance, on the one hand, internal energy tends to reach a minimum value, corresponding to maintaining e! order that the links present seek to impose, while, on the other, e! The corresponding TS mix energy term tends to a maximum value, corresponding to trying to maximize the disorder. For example, as the temperature of a solid such as copper increases, e! Disorder term is imposed on the effect of the bonds and this is how we can pass, under equilibrium conditions, from, ideally, a perfect crystal (T = 0 K) to a more imperfect URO, and then to a liquid and then to a steam It should be noted that for a strong bond solid, its internal energy E is approximately equal to its enthalpy H; This is due to the high intensity of their bonds, these materials have a low molar volume and only moderate changes in volume by varying P and T (within certain ranges).

Considering the previous two terms, the free energy (Gibbs) function is defined as G ^™ H-TS. Thus, the chemical equilibrium condition for a system with constant composition, temperature and pressure corresponds to a state in which such a system has a minimum of energy G; This is valid for all types of phases, not only solid. Gibbs' contributions to the development of equilibrium diagrams, as well as the importance of the latter, are well known to specialists. The emphasis of Gibbs' work was for the state we call stable equilibrium. On the other hand, the study of steels hardened by quenching in water and the discovery in 1908 of aging hardening of aluminum alloys (eg Ai-4.5% p. Cu), showed that numerous important systems are not strictly to balance, in the sense of corresponding to an absolute minimum of Gibbs free energy. This state is due to the fact that, at sufficiently low temperatures, the atomic movement (diffusion) in a solid becomes so slow that the atoms are frozen in their positions and the system is prevented from evolving. This kind of instant photography is currently considered a state of enormous importance in engineering and materials science. iíheím Ostwaíd (1853-1932), a professor of fa University of Leipzig, Nobel Prize in Chemistry (1909), was the first to clearly recognize this topic, introducing the concept of metastability. Figure 1 illustrates the concepts of a) metastable, b) unstable and c) stable status. In an unstable state the system evolves spontaneously with just a small disturbance. The determination of the stable and metastable equilibrium states concerns the equilibrium criterion (minimum values of the G function). Now the transformation of a subject! evolving from a metastable to a stable state, see Figure 1. Initially, the system must be removed from a well of initial potential Ga, raising its energy by ΔΘ ^* , by an external action, to reach the unstable intermediate state of energy Gb; and only after such a barrier has been overcome, the system will finally be able to arrive at the steady state of equilibrium associated with the energy well Ge. The net result is that the energy of the system has decreased by ΔΘ = Gc-Ga. In chemistry and materials, often the barrier ΔΘ * = Gb-Ga is overcome by the atomic vibrations associated with a sufficiently high temperature T (thermal activation). This barrier is called the activation energy of the transformation (or reaction) and its value is related to the intensity of the bonds and the atomic mechanism of the transformation involved. The speed at which this thermally activated transformation occurs is no longer a matter of the equilibrium criterion but of the "kinetics to reach equilibrium", which is strongly dependent on the values of AG ^* and T. frame, for a given solid one speaks of high, intermediate and low temperatures, depending on whether the transformation speed is high, moderate or practically zero; This is a simplification of a kinetics that varies continuously, exponentially, with temperature. There is also the transformation in which, from a metastable state, another metastable state is passed, but of lower energy G than the first.

Another notable example of classic metastable structure materials corresponds to window glass (SiQ ₂ ), an amorphous material {that is, with the structure of a supercooled liquid). Some examples of modern metallic materials of metastable structure are: materials manufactured by mechanical alloy (even incorporating ceramics) at nominal temperatures close to the environment; shape memory alloys (related to martensitic transformation); and solid amorphous alloys, today obtained at cooling rates as low as 1 K / s. Thus, the contributions of Gibbs and Ostwaid have been fundamental for the development of essential engineering tools and materials science, as are the phase diagrams a! balance and consideration of metastable states.

Thus, metastability is the property exhibited by a system with several equilibrium states, when it remains in a weakly stable equilibrium state during A considerable period of time. However, under the action of external disturbances (sometimes not easily detectable), these systems exhibit a temporary evolution towards a strongly stable state of equilibrium. Normally, metastability is due to slow state transformations. If we represent a physical-chemical system by its potential energy, a metastable state will be characterized by a state that corresponds to a local minimum of energy. In order for the system to reach the minimum energy state that corresponds to the thermodynamic equilibrium state, it is necessary to supply an amount of energy called activation energy.

For example in chemical metastability, at room temperature diamonds are metastable because the transformation to their stable form, graphite, is extremely slow. At higher temperatures and pressures, the transformation rate increases and graphite becomes diamond.

In a case of biological metastability, the links between Sos constructive elements of polymers such as DNA, RNA and proteins are also metastable. The carbon components that constitute living beings are metastable. An enzyme is a biocataiizer capable of establishing weak bonds with its substrate at the level of its active site. The bond energy released is at the origin of the decrease in the activation energy of the reaction. In this way, the reaction is activated kinetically (Arrhenius's law). An enzyme can accelerate the rate constant of the reaction it catalyzes: thus, very slow chemical reactions without a catalyst are accelerated until they can be used by the metabolism. In another aspect of physical metastability, a metastable state is a state that is a local minimum of energy, which is not fully stable under system disturbances above a certain magnitude. For example, in superfusion water, the drops of pure water in suspension in an also very pure air do not freeze at 0 ° C, but remain in a liquid state until reaching - 39 ° C. This state of overfusion ceases abruptly when the drop comes into contact with an external body (such as an ice crystal). On the other hand, for a radioactive isotope, the state of instability is characterized by the radioactive decay period, more or less long (ranging from minutes to several centuries and even millions of years). The Scottish physicist James Dewar (1842 - 1923) designed a vessel that bears his name - Dewar - to provide thermal insulation, reduce heat losses by conduction, convection or radiation, It is used to store liquids, cold or hot (Figure 2 ),

Its main use is in the storage of liquid nitrogen (whose boiling point is 77 K) and liquid oxygen (whose boiling point is 90 K), for a long time without the need for refrigeration. It can also be used to store helium, which has a very low boiling point, (4.2 K,) but the Dewar bottle must have a quadruple layer of glass and the space between walls must be filled with liquid nitrogen.

The type of construction of these containers makes them very sensitive when receiving blows and overpressures on the outside of the container. Thus, although the Dewar used as containers facilitate the transport and protection of materials that must be kept at a low temperature and protected from vibrations, agitation, goips, oscillations and / or any external disturbance that break their thermodynamic stability, these types of devices have some Common disadvantages such as: if the Dewar is dropped during handling, there is a risk that the glass will break; the cost of a traditional Dewar is high, for example, a 600 mi Dewar can have a sale price of more than $ 200; The classic Dewar is not necessarily easy to handle. However, there are other types of disadvantages in the use of Dewars for the transport and protection of samples, especially in the field of biomacromolecules. The Dewar have been frequently used for the transport and protection of crystalline compounds, including crystallized proteins that are subject to X-ray diffraction studies. However, during the transport route and mainly for chemical and biological safety reasons, they are reached to open the Dewar (for example, by customs agents) losing not only the low internal temperature of the container, but also the loss of intrinsic properties and / or decomposition of the material contained due to both the increase in temperature, and the incorrect handling of said material, such as protein degradation, which otherwise could not be transported under suitable conditions from one site to another, even on long journeys or intercontinental travel. There are other types of polymer containers that are molded from thermoplastic polymers, for example, polypropylene, polyethylene (including LLDPE, LDPE and, in particular HDPE), polyamide (Nylon), polystyrene, polyurethane, polyvinyl chloride, aceta! polyphenylene sulfide, polyesters, acryl-onitrile-butadiene-styrene (ABS) and other co-polymers of (aceta copolymers!) and the like, but which cannot be used since they do not prevent external disturbances, such as vibrations, agitation, shock or oscillations, or that affect thermodynamic stability, as well as changes in sample temperature.

US Patent 8,863,532 describes a system for use in the freezing, storage and defrosting of biopharmaceutical materials that includes a flexible sterile container with means for containing the biopharmaceutical material therein and a stiffer support than said container means. US Patent 5,344,036 describes a container system for reagents and other substances used in medical diagnostic devices. The container system includes a blow molded plastic vial, which can withstand lyophiization. For its part, US Patent 7,971, 744 describes a container for cryogenic separation and storage of fluids and fluid mixtures, made of cross-linked cross-linked polyolefsna foam with a density of between 2 and 4 inches per cubic foot. However, these types of containers are only to contain liquid N ₂ and none of the containers and devices of the previous documents are designed for the protection and transport of materials in which changes in temperature or external disturbances that break their stability as in the case of crystals, such as protein crystals.

The purpose of the present invention is to provide a new polyolefin device and method of transport and protection of materials against external disturbances and changes in temperature. The device of the present invention keeps the materials isolated from temperature changes, vibrations, agitation, shocks and / or oscillations, thus preventing their thermodynamic stability from breaking. SUMMARY OF THE INVENTION

It is an object of the present invention to provide devices or containers that allow the safe transport of samples or materials, which in addition to thermally insulating them, prevents external disturbances, such as vibrations, agitation, flutes or oscillations, affect the thermodynamic stability of the sample preventing it from losing its properties, mainly those that have to do with its solid state, such as crystals.

In another object of the invention, the materials are, without limiting the scope of the invention, selected from: biological macromolecules, proteins, crystallized proteins, crystalline compounds, co-crystals, polymorphs, hydrated polymorphs, solvated polymorphs, salts, powders, proteins in dust, and / or materials sensitive to changes in temperature. Said materials can preferably be found, for example, in the solid state, such as crystals. The crystals can be without being limiting of the invention, crystallized proteins.

It is another object of the present invention, a device comprising a container of plastic material, preferably polyolefin.

The device of the present invention comprises a container of plastic material, preferably of polyolefin, which has a series of perforations or holes drilled in a cylindrical shape in which a glass tube (401) is inserted cylindrically into which will deposit a material to protect. The glass tube (401) can also be, without limiting the scope of the invention, an elongated plastic or glass container, which in turn is a receptacle of at least one capillary tube (601), preferably of glass, which will contain the material to be protected and where the capillary tube (601) is optionally immersed or inserted in an insulating gel inside the glass tube (401). The glass tube (401) is selected from: nuclear magnetic resonance glass tubes, test tubes, blown glass tubes. It is preferably selected from tubes used in NMR.

The device of the present invention can be used to transport protein crystals safely, easily, without the need for N ₂ , without risks of tampering or external disturbances, without risks of thermal damage, and without the need to carry large and heavy loads. containers like a Dewar. In addition, its manufacture in plastic materials makes it safe and not dangerous, being able to even transport the sample by land, sea or area without risk.

In the following, the present invention, and particularly preferred embodiments thereof, will be described in greater detail in connection with the accompanying drawings. Figure 1. States a) metastable, b) unstable and c) stable, in a Gibbs free energy graph versus a variable that characterizes the progress of the process. The steady state of equilibrium and metastabie (s) correspond to minimums of the G function. The steady state of equilibrium is the absolute minimum.

Figure 2. View of a Dewar bottle or container.

Figure 3. Shows a front perspective view of the device for protection and transport, in which Figure 3A shows the device with housing holes (301) of cylindrical type in a container; Figure 3B shows the glass tube (401) of the type used in NMR together with its cap comprising at least one capillary tube (801), and Figure 3C shows the insertion of the glass tubes comprising said tube of glass in said housing and the means (201) for sealing or closing both said glass tubes each comprising at least one capillary tube (601), as well as said housings in the device container.

Figure 4, Means (201) for sealing or closing the housing holes of the container (301) and the cylindrical glass tubes (401) each comprising at least one capillary tube (601) in its bottom.

Figure 5. Infrared spectrum of two devices for protection of the present invention with different compactness.

DETAILED DESCRIPTION OF THE INVENTION

The purpose is to provide a new device for transport and protection of materials, such as biological macromolecules, against external disturbances and changes in temperature since keeping materials isolated from changes in temperature, vibration, shock or oscillation prevents breakage. Its thermodynamic stability. The device of the present invention allows transport in suitable conditions and without risks of tampering or external disturbances, and without risks of thermal damage, from one place to another, even on long journeys or intercontinental trips, materials that must be isolated from changes in temperature and / or external disturbances.

In the present invention, the term "material" or "materials" should be understood as those materials selected from biological macromolecules, proteins, crystallized proteins, crystalline compounds, co-crystals, polymorphs, polymorphs hydrates, solvated polymorphs, salts, powders, protein powders, and / or materials sensitive to changes in temperature. Said materials can preferably be found, for example, in a solid state such as crystals. The crystals can be without being limiting of the invention, crystallized proteins.

In the present invention, the term biological macromolecules should also be understood as proteins, nucleic acids and polssaccharides, and macromolecular complexes having combinations of these biomolecules.

At present, there are no devices that allow the safe transport of samples or materials, which in addition to thermally isolating them, prevent external disturbances, such as vibrations, agitation, shocks or oscillations, from affecting the thermodynamic stability of the sample preventing it from losing its properties, mainly those that have to do with its solid state, such as crystals.

The device according to the invention comprises:

»At least one container (101) of matter! polymeric and parallelepiped shaped;

"At least one housing hole (301) drilled in a cylindrical shape that is open at one end and closed at the other end, and said housing is arranged so that the open end is incorporated in said container and adjacent to the bottom thereof, and where the housing also allows to insert a cylindrical glass tube (401); and where two and up to three housing holes are preferably located for two and up to three cylindrical glass tubes in the container;

"A cylindrical glass tube (401) which in turn is a receptacle of at least one capillary tube (601), preferably of glass, into which a material or sample to be protected and transported will be deposited;

* means to seal or close the a! at least one, two and up to three holes for housing the container (301) and the at least one, two and up to three cylindrical glass tubes (401) comprising (n) each inside at least one capillary tube ( 801), wherein said means is a cover (201); and wherein said lid (201) optionally comprises a removable cap (501) for sealing or closing the at least one, two and up to three housing holes of the container (301) and the at least one, two and up to three tubes of cylindrical glass each comprising inside at least one capillary tube (601).

The container, of rectangular dimensions, comprises four walls and a bottom, where the open end of the container housing hole defines an inlet and e! closed end of the housing opening defines a closed end adjacent to the bottom of! container. The means (201), for sealing or closing the housing holes in the device container and the glass tubes each comprising inside at least one capillary tube (601), comprise perforations in its inner part {701} which fit perfectly with the container housing holes and with the cylindrical glass tubes (Figure 4),

In one embodiment, the entrance provides a housing hole for the sample or matter! In another embodiment, the inlet also provides a hole for the addition of an insulating gel, which flows freely throughout the housing of! Container of! device providing an additional protective layer if required, which ensures that thermal protection is effective.

In one embodiment, the perforated cylindrical shape of the housing (301) corresponds to the shape of a cylindrical tube, which can be a cylindrical glass tube (401) that is inserted through said container housing hole by its open end, and resting its base on the closed end adjacent to the bottom of! container. The glass tube (401) can be an elongated plastic or glass container, which in turn is a receptacle of at least one capillary tube (601), preferably of glass, which will contain the material! to protect. Said glass tube (401) is selected from among; nuclear magnetic resonance glass tubes, test tubes, blown glass tubes. It is preferably selected from glass tubes used in NMR.

In one embodiment, the cylindrical shape drilled from! housing (301) corresponds to the shape of a cylindrical tube, inside! which a glass tube (401) of the type used in NMR is inserted through said housing hole (301) of the container at its open end, and resting its base on the closed end adjacent to the bottom of the container. In turn, and optionally, the nuclear magnetic resonance glass tube (401) is closed with a removable plug (501), which is preferably a nuclear magnetic resonance tube stopper. A perspective perspective view of the device for protection and transport is shown in Figure 3, where Figure 3A shows the device with the cylindrical housing holes (301), with dimensions ranging from 4.5 long x 3.7 cm wide or deep x 8.0 cm high up to 75 cm long x 62 cm wide or deep x 8 cm high. According to the scope of the invention, and without limiting it, the device may comprise at least one container with 1 to N housing holes (301), wherein N is an integer between 1 and 50, preferably between 1- 25, and more preferably between 1-3, sufficient and capable of accommodating from 1 to N glass tubes, where N is an integer between 1 and 50, preferably between 1-25, and more preferably between 1-3. Preferably, the device has a dimension of 4.5 cm long x 3.7 wide or depth x 8.0 cm high, minimum and appropriate dimensions to accommodate from 1 to 3 glass tubes (401) each comprising inside a! minus a capillary tube (601). Figure 3B shows the glass tube (401), together with its cap, which will contain the material or sample to be protected; and wherein the tube (401) is selected from nuclear magnetic resonance glass tubes, test tubes, glass tubes, blown glass tubes; preferably the cylindrical glass tube is an NMR sample tube (401) together with its cap (501). In one embodiment, the glass tube (401) can also be, without limiting the scope of the invention, an elongated plasticized or glass container, which in turn is a receptacle of at least one capillary tube (801), preferably of glass , which will contain the material to be protected and where the capillary tube (801) is optionally immersed or inserted in a ge! insulator inside the glass tube (401). Figure 3C shows the insertion of the glass tubes comprising said glass tube in said housing and the means (201) for sealing or closing both said glass tubes each comprising at least one capillary tube (601), as well as said housings in the device container. Between the closed end of the housing and the bottom of the container there is at least a 1.0 cm gap or space. The means (201) for sealing or closing the housing holes of the device container and the cylindrical glass tubes, is a cover made of the same material as the rest of the device, with a dimension corresponding to the length and width of the container- of the device and a height of at least 1.0 cm and not more than 2.0 cm. In one embodiment, the means (201) for closing the orifice (s) of the container housing are also the means for closing the cylindrical glass tube (s). Optionally, the means (201) comprise a removable cap (501) for closing the container housing (s) and the cylindrical glass tubes. Preferably, when e! or the glass tubes (401), preferably a Nuclear Magnetic Resonance tubes, which in turn are a receptacle of at least one capillary tube (601) and up to N capillary tubes, where N is an integer greater than 1 that will contain the material to be protected, then said means (201) comprise removable caps (501) for nuclear magnetic resonance tube attached as a single piece or unit to said means for closing the container, its housings and the glass tubes comprising the capillary tube (s) (Figure 4).

In one embodiment, a glass tube (401), preferably a nuclear magnetic resonance glass tube, can comprise from one to N capillary tubes, where N is an integer greater than 1. The amount of capillary tubes (601) that They may be contained in a glass tube (401) will depend according to the specific storage and transportation requirements required of matter! or materials to be protected selected from among biological macromolecules, proteins, crystallized proteins, crystalline compounds, co-crystals, polymorphs, hydrated polymorphs, solvated polymorphs, salts, powders, powdered proteins, and / or materials sensitive to changes in temperature; and wherein said materials can preferably be found for example in the solid state such as crystals, as crystallized proteins; and wherein the biological macromolecules are selected from proteins, nucleic acids and polysaccharides, or macromolecular complexes that have combinations of these biomolecules.

The means for closing the orifice of the container housing or the glass tube comprising the capillary tube, or both at the same time, close and seal perfectly preventing the sample contained in the device from spilling, falling, saiga or draining. In an optional embodiment, preferably, a small cap (901) is inserted inside the cap (501) to avoid heat transfer in the sample or material to be protected (Figure 4). In one embodiment, the means (201) for closing e! or the hole (s) for housing the container (301), which may optionally also be the means for closing the glass tube in a cylindrical shape (401), are optionally attached to the container of! device through appropriate joining means (801) such as tape, tie, thread, flexible adherent materials and even but not limited to, a strip of polyolefin material which can be easily and quickly molded in a desired manner, such allowing for example the joining of the means to close the hole (s) of the container housing, with said hole container. Optionally, the device for protection and transport can be sealed or closed with adhesive tape usually used in electrical coatings.

The device container can be formed and sized according to the specific storage and transportation demands required. Preferably, the device container can be parallelepiped-shaped, like a rectangle. In an additional embodiment for example, the side walls of! container can be of equal dimensions essentially creating a square container. It will be understood thus, that the present invention is not limited and the device may comprise a container of any given size and may have substantially the same or similar size as other conventional cylindrical container, such as test tubes, glass tubes, tubes blown glass, plastic tubes, and / or plastic cylinders, preferably nuclear magnetic resonance tubes.

The device of the present invention comprises a container of polymeric material, preferably of polyolefin, which has a series of perforations or holes in which a cylindrical glass tube is inserted which in turn is a! less a capillary tube (601), preferably of glass, which will contain or inside! which will be deposited a material to protect. For example, when the material to be protected are protein crystals, they are grown in capillary tubes (801) with a diameter of 1mm. These are introduced into Sos tubes (401), which usually fit 4 to 5 glass capillary tubes 1mm in diameter. Once these have been introduced into the glass tube (401), it is understood that capillary tubes (801) of 1mm in diameter must be sealed on both sides, covered with a ge! Silica insulator, filling the tube (401) with a mixture of the components that will form the ge! insulating. This is an additional protective layer, which guarantees that thermal protection is effective. The tube (401) is selected from: nuclear magnetic resonance glass tubes, test tubes, glass tubes, blown glass tubes. It is preferably selected from tubes used in R N.

The device is made of a material! of polyolefin which can be molded easily and quickly in durable containers of a desired shape, such that they allow for example stacking.

The polyolefin material is a crosslinked and compact polyoiefin foam useful and suitable for transport and material protection devices, which in addition to allowing thermal insulation, prevents external disturbances, such as vibrations, agitation, shocks or oscillations that affect thermodynamic stability of the sample preventing it from losing its properties, mainly those that have to do with its solid state, such as crystals. Several characteristics of polyolefin foams make them advantageous for use in these devices such as: they are typically molded by conventional means well known to persons skilled in the art. Preferred methods include rotational molding, although it is contemplated that other methods may be used, such as injection molding, compression molding or extrusion and blow molding, and other conventional methods may also be used. Its cellular structure is typically thin enough to contain, for example, liquid nitrogen, without leaks; They are strong enough to withstand repeated exposures at a temperature scale ranging from cryogenesis to hot temperatures above 40 ° C. It is a non-reactive material; they have a low thermal conductivity and a low volumetric heat capacity, and also the polyolefin foam can withstand moderate physical manipulation and without mechanical failure, which makes this polyolefin material suitable and relevant for the manufacture of devices of the present invention. for transport and protection of materials such as biological macromolecules.

As used herein, the term "polyolefin foam" refers to a polyethylene foam, polypropylene foam, mixtures of polyethylene-polypropylene and copoimers, and foams containing a mixture or copolymer of olefin monomer and other monomers, in the extent to which mixed foams have at least some of the favorable characteristics indicated above. Besides of Polymers or polymers that make up the polyolefin foam, such as that of the present invention, may also have additives known in the art. For example, foams may include softeners, coloring agents, stabilizers, preservatives, and fillers.

Polyolefin foams are typically provided in a variety of densities. In the selection of the density of a polyolefin foam to be used for a device such as that of the present invention, one must seek to balance the mechanical and thermal properties of the material. If the foam is not very dense, it cannot have advantageous mechanical properties; If the foam is too dense, it will have a higher thermal conductivity. In the present invention, it has been found that the polyolefin foam has a density of at least 32.04 kg / m ³ (2 pounds per cubic foot), while the densities in the range of 32.04 to 64.07 kg / m ³ (2 pounds at 4 pounds per cubic foot) may be particularly advantageous in the present invention. However, in some embodiments of the invention, polyolefin foams are expected to comprise densities of 98.11-128 kg / m ³ (6-8 pounds per cubic foot).

The device described in the present invention is capable of withstanding temperature variations and even withstanding temperatures up to -160 ° C without presenting temperature variations.

The device or container for transporting crystallized proteins, whose crystalline phase is easily lost due to changes in temperature and / or external disturbances, has been successfully tested. For example, liasas constitute a diverse group of water-soluble enzymes that catalyze the hydrolysis of ester bonds in water-insoluble lipid substrates. Iipases play essential roles in the digestion, transport, and treatment of dietary lipids such as triacylglycerides, fats, oils in most, if not all, of living organisms. Likewise, üpasa B is industrially important in the synthesis of glycolipids. Lipase B in particular is a pure, dry, crystalline form of üpase B from Candida Antarctica produced by submerged fermentation of a genetically modified microorganism of Aspergiiius oryzae. Lipase B is a white crystalline powder, where lipase B is present in the form of crystals and there are no other ingredients or buffer salts. Lipase B has a pH of 5-7, and has a isoelectric point of 6.0. AND! Mr of Lipase B is 35 kDa by SDS polyacrylamide gel electrophoresis,

In the present invention, lipase has been used as a thermal sensor since the crystals are sensitive to dissolve when there are changes to their crystallization temperature that is 18 ° C and variations in a range of ± 2 ° C that affect it considerably. In intercontinental routes and with changes in atmospheric pressure and temperature, it has been possible to verify that the lipase remains in the crystalline phase without changes, which guarantees that any other sample of a crystal to be analyzed - and of which it is unknown if the temperature changes and / or disturbances will affect its thermodynamic stability - it remains intact without modification of its physicochemical properties, including its crystalline phase.

In parallel to this experiment, 2 proteins (glucose isomerase and iisozyme) have been transported to two different synchrotons each located in Trieste, Italy (Elettra) and SLAC in Stanford, California, USA. In both cases both crystals (transported in this device) diffracted the X-rays at a very high resolution, which meant that there was no structural damage in the transport. As an example, the following tables 1 and 2 are presented with the results:

Table 1. Glucose isomerase.

* Without influence of magnetic field in its growth.

Crystal 1 and Crystal! 2 were grown in the presence of a magnetic field of

Table 2. Lysozyme

* Without influence of magnetic field in its growth.

Crystal 1 of Table 2 was grown in the presence of a 700 MHz magnetic field.

When talking about 1A resolution, it means that it is possible to appreciate the electronic densities clearly, and therefore the transport did not affect the biomachromolecules in its crystallographic structure. EXAMPLES

Example 1. Manufacturing of the device

Pieces of the polyolefin of the following dimensions are cut: 4.5 cm long x 3.7 wide or depth x 8.0 cm high, minimum dimensions and appropriate for Ajar from 1 to 3 tubes (401). Once the cuts are made, a parallelepiped shaped device is obtained. Subsequently, 3 perforations are made with a drill the size of the NMR tubes. Between the closed end of the housing and the bottom of the container there is at least a 1.0 cm gap or gap. The means to close the device container is a lid (201) made of the same material as e! rest of the device, with a dimension corresponding to the length and width of the device container and a height of at least 1.0 cm and not more than 2.0 cm. Once the perforations have been made, Sos crystals of proteins to be protected thermally and against disturbances, they are grown in capillary tubes (601) with a diameter of 1mm. These are introduced into the RN tubes (401), which usually fit 4 to 5 glass capillary tubes of 1 mm in diameter. Once introduced these in e! NMR tube, it is understood that the 1mm diameter capillary tubes must be sealed on both sides with plasticine or wax (to prevent the proteins contained in them from dehydrating and leaking the mother liquor), then covered with a gel Silica insulator, filling the NMR tube (401) with a mixture of the components that will form the gel (1 mL of 1 M acetic acid with two mL of sodium metasilicate density of 1.08 g / mL). This is an additional protective layer, which ensures that thermal and external disturbance protection is effective. The NMR tube (401) containing the 1mm diameter capillary tubes is covered or closed with the lid (201) which is a cover of the same material of the device (polyolefin), which also comprises a removable cap (501) than in its interior optionally comprises a small plug (901) (Figure 4); optionally the lid (201) is attached to the device container through appropriate joining means (801) such as tape, tie, thread, flexible adhesive materials and even but not limited to, a strip of polyolefin material which it can be molded easily and quickly in a desired manner; optionally, the device for protection and transport can be sealed or closed with adhesive tape usually used in electrical coatings. Example 2. Infrared spectrum of two devices for protection of the present invention with different compactness.

An infrared spectrum of two devices for protection of different compaction is shown in Figure 3. The red band (A) is a device of material compacted and preferred in the invention. In black (B) it is a device of less compacted material but equally efficient in the invention.

In both cases, bands are observed in the region between 2750-3000, 1500 and 750 cm -1, The difference in% of transmittance is precisely due to! degree of compaction that each device has.

Example 3. Insulating gel

As defined above, the device of the present invention comprises a container of plastic material, preferably of polyolefin, which has a series of perforations in which a cylindrical glass tube is inserted into which a material to be protected will be deposited. , preferably, the material to be protected is in turn inside a capillary tube (801). Thus, the glass tube (401) can be, without limiting the scope of the invention, a nuclear magnetic resonance tube, which in turn is a receptacle of a capillary tube that will contain the material to be protected. The material to be protected, for example a biomachromolecule, can be placed, placed or even crystallized in situ in said capillaries which will be introduced into the tube (401) (for example, an NMR tube) as described in Example 1 In turn, a gel is used in which these capillaries will be immersed or inserted. Said gel allows to completely isolate external disturbances such as vibrations, agitation, blows and / or oscillations, as well as temperature changes, to the sample of matter! or materials transported in the device. The manufacture of said gel comprises the following steps and compounds:

A solution of sodium metasilicate of a density = 1.06 g / ml is taken and neutralized with a solution of 1 M acetic acid. The proportions to produce a gel at ρΗ ^ 7 are:

- 1 ml of acetic acid and 2 ml of the above sodium metasilicate solution are initially added to the mixture; Y

- the gelation proceeds in an approximate time between 3 and 5 minutes.

Example 4, transport and protection of materials with the device of the present invention. As mentioned above, the protein crystals grown in capillary tubes (601) of 1mm in diameter, are introduced into the NMR tube (401) and the components that will form the silica gel are added. It is sealed e! device. The crystals can then be transported in a portfolio and there is no need to worry that they suffer any damage due to thermal effects (sudden changes in temperature) or other disturbances such as oscillations, vibrations, agitations or blows. When you arrive at the site where the X-ray data will be collected (either a synchrotron, for example) or just some distant laboratory, with which you have some collaboration, for e! Crystal analysis, these are removed from the device. The capillaries of i mm in diameter are removed with fine-tip tweezers from the NMR tube and cut from both sides (these were sealed with plastiiin or wax) to be able to extract the protein crystals. Tables 1 and 2 show e! As a result of having transported two model proteins, which were grown in the presence of very intense magnetic fields, the resolution data is between 1.5 Á and 1.1 This, in crystallography, implies that these crystals are of excellent quality. These were transported from Mexico City to the Elettra Synchrotron in Triste (Italy) in March 2016 and a second consignment was subsequently transported to the Stanford California Synchrotron (SLAC) in the US at the end of April 2016. These two Experiments perfectly validate the effectiveness of the device that is intended to be patented.

Industries Application

The device of the present invention can be used to transport protein crystals safely, easily, without the need for, without risks of tampering or external disturbances, without risks of thermal damage, and without the need to load large and heavy containers. Like a Dewar In addition, its manufacture in plastic materials makes it safe and not dangerous, being able to even transport the sample by land, sea or area without risk.

The invention can be applied - without being limiting in its scope - in the protection and transport in general of any material that must be isolated from changes in temperature and / or external disturbances. A very useful application is for those people who use synchrotons, who for their work and sophisticated work equipment required for e! study of material samples, selected from biological macromolecules, proteins, crystallized proteins, crystalline compounds, co-crystals, polymorphs, hydrated polymorphs, solvated polymorphs, salts, powders, powdered proteins, and / or temperature sensitive materials - preferably crystallized proteins and / or powdered proteins - they travel to institutions in different parts of the world that have this kind of equipment. Thus, the present invention provides a polyolefin device and method of transport and protection of this class of materials that must be kept isolated from changes in temperature, vibrations, agitation, shocks and / or oscillations, thus preventing its thermodynamic stability from breaking, thus allowing to keep intact the material to be subjected to tests, for example, of X-ray diffraction or other crystallographic studies.

The methods and techniques illustrated in the previous construction examples are advantageous because they allow the device for protection and transport to be formed and sized according to the specific storage and transport demands required. Also, those skilled in the art will realize that the polyolefin material can be cut in the form of a storage vessel by conventional molding, blow molding or other techniques,

Although the invention has been described with respect to certain embodiments to be implemented, the description is intended to be by way of example, rather than limiting. Modifications and changes can be made within the scope of the invention, as set forth in the following claims.

Claims

! - A device for transport and protection of materials against external disturbances and changes in temperature, characterized in that it comprises:

· At least one container (101) of poSimeric material and in the form of a parallelepiped;

• at least one housing hole (301) drilled in a cylindrical shape that is open at one end and closed at the other end, and said housing is arranged such that the open end is incorporated in said container and adjacent to the bottom thereof, and where the housing also allows to insert a cylindrical glass tube (401); and where two and up to three housing holes are preferably located for two and up to three cylindrical glass tubes in the container;

»A cylindrical glass tube (401) which in turn is a receptacle of at least one capillary tube (801) of 1 mm in diameter, preferably of glass, into which a material or sample to be protected and transported will be deposited;

_* means for sealing or closing the at least one, two and up to three container housing holes (301) and the a! minus one, two and up to three cylindrical glass tubes (401) that each one comprises (n) inside it at least one capillary tube (801), wherein said means is a lid (201); and wherein said lid (201) optionally comprises a removable plug (501) for sealing or closing the at least one, two and up to three holes for housing the container (301) and the at least one, two and up to three glass tubes cylindrical ones each comprising at least one capillary tube (601); and wherein the removable cap (501) optionally comprises a small plug (901) inside.

2.- E! device according to claim 1, characterized in that the means (201), for sealing or closing the housing holes in the device container and the glass tubes (401) comprising (n) each inside therein at least one capillary tube (801), comprise perforations in its internal part (701) that perfectly assemble with the holes of the container and with the cylindrical glass tubes.

3. The device according to claim 1, characterized in that the glass tube {401} which in turn is a receptacle of at least one capillary tube can contain up to 5 capillary tubes (801) of 1mm diameter glass which will contain (n) the material to be protected,

4. The device according to claim 3, characterized in that the glass tube (401) is selected from: nuclear magnetic resonance glass tubes, test tubes, blown glass tubes,

5. The device according to claim 4, characterized in that the glass tube (401) is preferably an NMR glass tube with a detachable cap (501) for nuclear magnetic resonance tube which optionally comprises a small inside stopper (901).

8. The device according to claim 1, characterized in that the means (201) for closing the orifice (s) of housing of the container are also the means for closing the glass tube cylindrically,

7. The device according to claim 1, characterized in that the means (201) for closing the housing holes of the container (301), optionally are connected to the device container through joining means (801),

8. - The device according to claim 7, characterized in that the joining means are selected from: tape, bond, thread, flexible adhesive materials, or a strip of polyolefin material,

9. - The device according to any of the preceding claims, characterized in that the means for closing the hole of the container housing (301) or the glass tube (401) comprising at least one capillary tube (801) inside , or both at the same time, close and seal perfectly preventing the sample contained in the device from spilling, falling, leaving or draining.

10. - The device according to claim 1, characterized in that the polymeric material is a polyolefin.

11. - The device according to claim 1, characterized in that the material to be transported and protected is selected from among biological macromolecules, proteins, crystallized proteins, crystalline compounds, co-crystals, polymorphs, hydrated polymorphs, solvated polymorphs, salts, powders, proteins powder, and / or materials sensitive to changes in temperature.

12. - The device according to claim 1 characterized in that the biological macromolecules can be proteins, nucleic acids, polysaccharides, or macromolecular complexes having combinations thereof.

13. - The device according to any of claims 11-12, characterized in that the material is preferably crystallized proteins.

14. - Use of the device of claim 1 for the safe transport and protection of materials against external disturbances and changes in temperature.

15. - E! Use of claim 14 wherein the materials are selected from or among biological macromolecules, proteins, crystallized proteins, crystalline compounds, co-crystals, polymorphs, hydrated polymorphs, solvated polymorphs, salts, powders, powdered proteins, and / or materials sensitive to changes in temperature.

16. - The use of claim 15 wherein the material is preferably 5 crystallized proteins.

17. - The use of claim 14 wherein the external disturbances can be vibrations, agitation, blows or oscillations that affect the thermodynamic stability of the material. 0

5

0