KR100473538B1 - Irradiation device and method for fluids especially for body fluids - Google Patents

Abstract

The present invention relates to an apparatus 1 suitable for use in the sterilization of a fluid 17, such as a biological fluid containing lymphocytes and / or microorganisms or fractions thereof. The device 1 comprises a container 2 with an inlet 4 and an outlet 5 and a passage 3 extending linearly between the inlet and the outlet, and a heat exchange device 14. The passage 3 has a heat exchange surface 10 which is in direct contact with the heat exchange device 14 so that heat passes therethrough. The temperature control means 7 is arranged and arranged to keep the temperature of the fluid 17 in the passage 3 lower than the temperature at which the components of the fluid can form insoluble particles during irradiation. The passage 3 has walls that penetrate lymphocytes and / or microbial inactivating radiation 20. The passage 3 is the heat exchange face in the irradiation zone 19 in which the entirety of the fluid 17 passing between the inlet 4 and the outlet 5 in use of the device extends along the wall. A static mixer 6 is arranged that is configured and arranged to thoroughly mix the fluid 17 to make contact with the 10. Thereby, in use of the device 1, the entirety of the fluid 17 passing through the container 2 can be exposed to irradiation at an equal level while maintaining a safe temperature.

Description

Irradiation device and method for fluids especially for body fluids

The present invention is intended for use in treating biological fluids, in particular fluids thereof and selected components of fractions thereof, for example to inactivate lymphocytes, microorganisms, viruses and the like in human blood. A device suitable for the present invention.

Large amounts of body fluids, such as blood and plasma, and various fractions thereof are used to treat patients suffering from various conditions. However, contamination of such fluids with various viruses and other microorganisms can cause serious new conditions in patients receiving these fluids and may even lead to death.

It is known that ultraviolet (UV) irradiation can inactivate lymphocytes and viruses, but this has not been practical because the UV transmittance of blood is so low that it is difficult to ensure complete irradiation and inactivation. Recently, the inventors have significantly reduced this problem by using a static mixer in British Patent No. 2,200,020 which allows the mixture of the fluid to be mixed very thoroughly during ultraviolet irradiation so that the entire fluid can be irradiated substantially uniformly. .

The inventors have found that fluids containing fibrinogen are easy to activate and tend to form rather large polymeric fibrin particles. In addition, such particles can form around viruses and other microorganisms, shielding them from UV radiation, thereby preventing their inactivation and seriously endangering the health of recipients of the treated fluid. Such fibrinogen activation can be easily caused by mechanical stresses, such as, for example, shear forces present upon mixing and heat that can easily occur locally during irradiation. Insoluble particle materials may also be formed by thermal and / or mechanical modification of other proteinaceous components.

In addition, the above problem also occurs in the conventional sterilization of human blood products, which generally involves long-term warming of approximately 48 to 72 hours at a temperature of about 78 ° C. This treatment also has the disadvantage of being relatively time consuming and taking up the majority of relatively large devices, and can result in a significant loss of the effectiveness of the blood product.

1 is a partial cross-sectional schematic diagram of a device according to the invention.

It is an object of the present invention to avoid or minimize one or more of the above disadvantages.

The inventors have found that by carefully controlling the temperature of the fluid and preventing local heating of the fluid, it is possible to prevent the formation of particles that can shield viruses and other microorganisms from inactivating radiation.

According to one aspect of the present invention there is provided a device for use in sterilizing a biological fluid containing lymphocytes and / or microorganisms or a fraction thereof, the apparatus comprising a passage extending linearly between the inlet and the outlet and the inlet and the outlet. A container having a means, a heat exchanger in direct contact with heat exchange surfaces passing through the passage means, and a temperature of the fluid in the passage means for insoluble particles during irradiation. And a temperature control means constructed and arranged to maintain a temperature below a temperature that can be formed, said passage means having walls for transmitting lymphocytes and / or microbial inactivating radiation, and furthermore, in use of said device, said inlet The heat exchange surface in an irradiation zone in which an entirety of the fluid passing between the outlet and the outlet extends along the wall And a static mixer configured and arranged to thoroughly mix the fluid so that, in use of the device, the entirety of the fluid passing through the vessel can be exposed to irradiation at an equal level while maintaining a safe temperature. A biological fluid sterilization apparatus is provided which is housed in a passage means.

By using the device of the present invention, a very uniform treatment of the fluid can be achieved with respect to both irradiation and temperature, so as to be surrounded by insoluble material formed by shielding by excessive depth of the soluble fluid component or by somewhat direct thermal denaturation of the fluid component. Lack of exposure to inactivated radiation as a result of shielding by or as a result of thermal and / or mechanically induced reactions (such as fibrinogen activation), as well as reducing and / or degrading the inactivation effect ( Local overheating by irradiation that can lead to an increase in gasification can be avoided, thus maximizing the inactivation of lymphocytes (as needed) and / or undesirable microorganisms, while denaturing and degrading useful fluid components. Minimization can be achieved.

Various types of heat exchangers can be used, including solid state heat exchangers such as the Peltier effect heat exchanger. Preferably, a heat exchanger device is used in which heat exchange fluid (eg, a gas, a liquid, or a liquid mixed with a gas and / or a frozen liquid) is circulated therein. This is because it generally facilitates more precise control of biological fluid temperature.

It is preferable to use a ring-shaped container having an outer wall which substantially transmits lymphocytes and / or microbial inactivating radiation and an inner wall constituting the heat exchange surface. The inner wall is preferably made of a high thermal conductivity, generally inert, physiologically compatible material, such as, for example, stainless steel. Any suitable heat exchange fluid may be used, such as, for example, water. If a heat exchange fluid is used, the temperature of the fluid can be controlled in various ways away from the heat exchange face in the vessel, for example using a solid heat exchanger such as a Peltier effect heat pump or cooling coil or the like.

Various types of temperature control means can be used. Generally, a cooling rate having a fluid passage means, a heat exchange surface, and a controller for changing the cooling rate in response to an input from a temperature sensor means arranged in contact with heat to pass through at least one of the heat exchange fluid passages in the heat exchange device. Variable chillers are used.

The "safe" temperature range for avoiding denaturation and / or other heat-induced reactions may vary depending on the biological fluid or fraction thereof, and also depending on the use of the fluid, and the fluid temperature may be subject to many different values as desired. It will be appreciated that a temperature control means is used which allows it to be maintained at. In general, for body fluids containing fibrinogen, the temperature is suitably maintained at 37 ° C. or lower, preferably at −5 to 37 ° C., advantageously at −5 to 19 ° C.

In a preferred embodiment of the present invention, the device is used such that the expression transparent wall (transparent wall is merely used to indicate substantial transmission of inactivated radiation, for example, important transmission of other wavelengths such as visible light). Or at least one lymphocyte and / or microbial inactivating radiation source, mounted somewhat in proximity to (or may not be accompanied by). The mounting location of this radiation source is generally defined to minimize the undesired heating of the transparent wall and the biological fluid in contact with it while maximizing the radiation intensity in the irradiation zone. Depending on the radiation source used, the radiation source is usually placed 1.5 to 5 mm away from the transparent wall.

Various lymphocytes and / or microbial inactivated radiation can be used, but in general ultraviolet (UV) is preferred. In particular, UV in the wavelength range of 100-400 nm, preferably 200-350 nm, for example UVA of approximately 320-400 nm, UVB of approximately 310 nm, and UVC of approximately 254 nm are preferred.

Suitable UV sources are readily available commercially. Specific UV sources include GTE Sylvania Ltd., West Yorkshire, Schiphol, Charlestown, UK. Products include Middlesex, UK, Thorn EMI from Enfield, Sherlay, UK and Philips Lighting from Croydon.

Also within the scope of the present invention are microbial indirect inactivation wherein a photoactive agent that is converted from inactive form to lymphocyte and / or microbial inactivation form by UV irradiation is introduced into the fluid. One example of this type of photoactive agent is psoralen, for example 8-methoxy sorenene, which is capable of forming photoadducts with DNA in lymphocytes when exposed to UVA at 320-400 nm wavelength. Can be mentioned.

When UV radiation is used to effectuate inactivation, the sidewalls of the container may be, for example, silica and other ultraviolet transmissive glass (sold under the trade names Spectrosil and Vitreosil); silicon; Cellulose products such as Cellophane (trade name); And plastic materials such as polytetrafluoroethylene (PTFE), fluorinated ethylene propylene (FEP) and preferably low density polyethylene (LDPE) or polyvinyl chloride (PVC).

Other inactivating radiations that may be used include, for example, microwave lines used in connection with glass or ceramic vessel walls; Infrared light used, for example, in connection with quartz container walls; For example, ultrasonic lines used in connection with stainless steel vessel walls are mentioned.

The required irradiation time depends on the intensity, placement and number of sources used, the permeability of the container sidewall material, the container shape, the mixing efficiency within the container, the surface area of the thin fluid layer adjacent to the container sidewall, and the container. It depends on various factors such as the length of the passage means within, the flow rate of the fluid being treated, the residence time of the fluid in the irradiation zone, and the nature of the fluid itself. However, the irradiation time required can be readily determined by simple trial and error using appropriate techniques known in the art to assess the inactivation of the relevant microorganisms, and more details are described below. In general, the residence time in the vessel is between 5 seconds and 30 minutes, preferably between 30 seconds and 10 minutes, for example 2 minutes, and the vessel sidewall material and the thickness and the radiation source are effective within such time. Selected and arranged to provide a line.

In addition, the required irradiation time can be achieved by a number of different methods, including one or more of the methods listed below. That is, the method may include using vessels with irradiation zones of different lengths, varying the flow rate of the fluid, using multiple devices in series, and circulating the fluid multiple times through the device (s). In general, however, in particular when the inactivation treatment system is integrated into a production line for the production of various products, for example IgG, factor VIII, etc., the level or irradiation required in a single passage is achieved. It is highly desirable that an inactivation treatment system be designed.

While it is a particular advantage of the present invention that minimization of useful fluid components is minimized, it may be advantageous to include one or more protective agents in the body fluid that further reduce denaturation or degradation of the useful fluid components. Such protective agents include rutin, ascorbic acid (ca 1 mM) and quercetin (ca 0.2 mM).

In addition, the degree of mixing required to achieve complete irradiation depends on various factors such as the permeability of the fluid to inert radiation and the total depth of the fluid in the vessel from the wall through which the radiation penetrates. In general, the lower the permeability and the greater the fluid depth, the greater the number of mixer elements and the number of mixing stages required.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings.

1 shows an apparatus 1 according to the invention, which has an inlet 4 and an outlet 5 and comprises a container 2 in the form of a cylindrical tube 3 of quartz or other ultraviolet (UV) transmissive material. The apparatus further includes a static mixer 6 and a temperature control means 7 extending in the axial direction. More specifically, the static mixer 6 comprises a series of spiral “screw” members 8 extending axially and angularly offset, the screw members 8 being evenly separated. However, it forms a plurality of pairs of flow paths that are coalesced at confluence points 9 between successive screw members 8, thus providing an exponentially increasing degree of mixing depending on the number of screw members used. do.

The screw members 8 are mounted on a hollow conduit 10 which forms a heat exchange fluid passage 11 forming part of the temperature control means 7. More specifically, the temperature control means 7 comprises a heat exchange circuit 12 having a pump means 13 for circulating the heat exchange fluid 12a and a Peltier effect heat exchanger 14. . The heat exchange device 14 is provided with a control means 15 having a temperature sensor 16 installed in the vessel 2 for monitoring the temperature of the fluid under irradiation. The control means 15 are configured and arranged to control the cooling rate provided to maintain the desired fluid temperature. This temperature may be a fixed value, or more conveniently, the control means 15 may be provided with user operable input means for changing the desired temperature setting.

The conduit 10 constituting the heat exchange surface (preferably also with the screw member 8) facilitates effective heat transfer between the fluid 17 to be treated and the heat exchange circuit 12 to within a relatively narrow range. By tightly controlling the fluid temperature, on the one hand, inerts such as stainless steel can be used to maximize the efficiency of the sterilization / inactivation treatment and, on the other hand, to minimize undesirable denaturation or degradation of useful fluid components. And is made of a physiologically acceptable thermally conductive material.

Irradiation of the radiation 20 is carried out by a plurality of UVC emitting fluorescent tubes 21 which are spaced apart from the vessel 2 at close intervals and extend parallel to the vessel and are distributed at an angle around the vessel. . It is preferred that a reflector 22 is provided to focus the radiation 20 on the container. In addition, the irradiation chamber may be cooled by a fan 23. The vessel 2 is made of quartz and has a diameter of approximately 20 mm in order to maximize the transmission of radiation 20 into the fluid 17 being processed, the static mixer 6 has a length of 300 mm, and 10 screws Has a member.

The pump means 13 deforms or degrades the useful fluid component due to poor fluid residence time in the vessel 2 in the irradiation zone 19 and due to mechanical stress in the static mixer 6 and its surroundings. It is desirable to have a flow rate controller for changing the flow rate of the heat exchange fluid 12a in order to minimize. In general, a flow rate of 1 cm / sec to 100 cm / sec, preferably 2 cm / sec to 50 cm / sec, more preferably 5 cm / sec to 20 cm / sec may be used.

It will be appreciated that the vessel 2 and the static mixer 6 can be constructed and arranged such that complete irradiation can be achieved in one pass of fluid through the vessel. However, multiple passes may be used to achieve sufficient irradiation.

The use of this apparatus is further described in the following examples.

Example 1 Treatment of Human Plasma

Irradiation device with four 500 mm long UVC light sources arranged around a 6 mm inner diameter PTFE tube containing a 34 cm long static mixer of the type shown in FIG. 1 with 48 screw members. Irradiation was carried out using. The fluid was circulated through a quartz tube and a cooling device installed in series with the quartz tube at a flow rate of 100 ml / min. This flow rate corresponds to the irradiation time of about 6.2 seconds in one passage. The fluid was circulated until a total effective irradiation time of approximately 100 seconds was achieved and its temperature maintained at approximately 6.5 ° C.

Plasma samples (200 ml) into which bacteriophage virus (2 ml) selected from X174 and MS-2 (single chain DNA and RNA, respectively), T4 (double chain DNA) and PR7772 (enclosed double chain DNA) were introduced. When used, virus kills in the 5-6 log range (ie, over 99.999%) were achieved.

At the same time, the coagulation factor activity of the major components of plasma remained substantially as follows.

Factor CIII: C 57.3 ± 4.2%

Factor V 38.8 ± 10.4%

Fibrinogen 63.5 ± 4.2%

APTT 18.5 ± 3.4%

In another experiment, rutin (1.6 mM) was introduced into the plasma as a protective agent and the retention of coagulation factor activity was increased above 85% while maintaining virus inactivation levels.

Example 2

Using a substantially similar procedure, an irradiation time of 300 seconds was performed on a human immunoglobulin preparation (150 g IgG per liter) containing MS-2 (1.5 g).

Aggregation increased only from the initial level of 7.0% to 7.6%, but a level of virus inactivation of 4.8 logs was achieved.

Sterilization of blood was monitored by one or more of the following procedures.

(a) Lymphocytes are isolated and cultured, then tritiated thymidine is administered and liquid scintillation is counted.

(b) Lymphocytes are isolated, cultured and examined under an electron microscope.

(c) Isolate lymphocytes and observe the response to tissue dyes.

(d) Cultivate the bacteria by standard experimental methods.

(e) Incubate the virus by standard experimental methods.

(f) Protozoa are studied by optical and electron microscopy and by in vivo passage of animal species.

(g) Study the biological trends of platelets, such as reactions in agregometers and post-exposure to collagen, ATP, etc. by standard hematological in vitro techniques.

Claims (16)

A device used to sterilize any of lymphocytes and microorganisms or fluid 17, which is a biological fluid containing fractions of lymphocytes and microorganisms, or a fraction thereof, wherein the apparatus 1 comprises an inlet 4 and an outlet 5 ) And a vessel (2) having tubular passage means (3) extending linearly between the inlet and the outlet thereof, the heat exchange surface (10) passing through the passage means (3) and the heat directly therethrough. Configured to keep the temperature of the heat exchanger 14 in contact and the fluid 17 in the passage means 3 below a temperature at which the components of the fluid 17 can form insoluble particles during irradiation. And a temperature control means (7) arranged and arranged, the passage means (3) has a wall for transmitting radiation (20) for inactivating any of lymphocytes and microorganisms or both lymphocytes and microorganisms, The passage means (3), With the heat exchange face 10 in the irradiation zone 19, the entirety of the fluid 17 passing between the inlet 4 and the outlet 5 in the use of the device 1 extends along the wall. In contact with the fluid 17 to ensure that the entirety of the fluid 17 passing through the container 2 during use of the device 1 is exposed to irradiation at an equal level while maintaining a safe temperature. A biological fluid sterilization device characterized in that it contains a static mixer (6) arranged and arranged to mix. 2. A biological fluid sterilization apparatus according to claim 1, wherein the heat exchanger is a Peltier effect heat exchanger. The heat exchange surface of claim 1, wherein the heat exchange surface comprises a conduit (10) through which heat exchange fluid (12a) passes inside the static mixer (6), and the conduit passes through heat exchange surface and heat of the heat exchange device (14). Biological fluid sterilization device, characterized in that in direct contact with. The heat exchange surface (10) and the outer wall (1) according to any one of claims 1 to 3, wherein the vessel (2) transmits an outer wall through which radiation (20) inactivating either lymphocytes or microorganisms or both lymphocytes and microorganisms is inactivated. Biological fluid sterilization apparatus characterized in that the ring shape having an inner wall constituting. 4. A biological fluid according to claim 3, characterized in that the temperature of the heat exchange fluid 12a is controlled by a solid state heat exchanger 14 at a distance from the heat exchange face 10 in the vessel 2. Sterilization device. The heat control means according to claim 3, wherein heat is passed through at least one of the passage means (3), the heat exchange surface (10), and the heat exchange fluid passage (11) in the heat exchange device (14). so that a cooling rate of the biological fluid sterilizing device, characterized in that the form of adjustable cooling unit includes a controller 15 for varying the cooling rate according to an input from the temperature sensor 16 it is placed in contact. 4. The temperature control means (7) according to any one of claims 1 to 3, wherein the temperature control means (7) sets the temperature of the body fluid to -5 ° C to +37 for use in sterilizing body fluids containing fibrinogen as the biological fluid. Biological fluid sterilization device, characterized in that it is configured and arranged to maintain a temperature in the range of ℃. 4. The passage means (3) according to any one of claims 1 to 3, wherein at least one radiation source (21) which emits the radiation which inactivates either lymphocytes or microorganisms or both lymphocytes and microorganisms. Biological fluid sterilization apparatus, wherein the biological fluid sterilization apparatus is spaced apart from each other in close proximity to the wall. 4. A biological fluid sterilization apparatus according to any one of claims 1 to 3, wherein said radiation for inactivating any of lymphocytes and microorganisms or both lymphocytes and microorganisms is ultraviolet light having a wavelength in the range of 200 to 350 nm. . The biological material according to any one of claims 1 to 3, characterized in that the side wall of the container (2) is made of an ultraviolet transmitting material selected from the group consisting of ultraviolet transmitting glass, silicone, cellulose products and plastic materials. Fluid sterilization device. The apparatus of claim 1, further comprising: microwave lines used in connection with glass; Infrared light used in connection with quartz container walls; A biological fluid sterilization device characterized in that a combination of inert radiation and container wall material selected from the group consisting of ultrasonic lines used in connection with metal container walls is used. 4. The apparatus of claim 1, further comprising reflectors 22 spaced apart around the vessel 2 to focus the radiation 20 on the vessel 2. Biological fluid sterilization apparatus, characterized in that. A method of sterilizing a biological fluid 17 or a fraction thereof containing either lymphocytes and microorganisms or both lymphocytes and microorganisms, Providing an apparatus (1) according to claim 1; At a location proximate the transparent wall of the device (1), providing a radiation source (21) which emits radiation which inactivates either lymphocytes or microorganisms or both lymphocytes and microorganisms; Passing the fluid 17 through the passage means 3 of the device 1 such that the entirety of the fluid 17 is exposed to an equivalent level of either lymphocytes or microorganisms or radiation inactivating both lymphocytes and microorganisms. step; And Temperature control means of the apparatus 1 to maintain the temperature of the fluid 17 in the passage means 3 below a temperature at which components of the fluid 17 can form insoluble particles during irradiation ( 7) operating the biological fluid sterilization method comprising the step of operating. 14. The fluid 17 to be sterilized, according to claim 13, prior to said step of passing the fluid 17 through the passage means 3, to the fluid 17 to be sterilized, either lymphocytes and microorganisms or both lymphocytes and microorganisms from the inactivated form by radiation. A biological fluid sterilization method further comprising the step of introducing a photoactive agent that is convertible into an inactivated form. 15. The fluid 17 according to claim 13 or 14, in order to further reduce denaturation or deterioration of useful fluid components, prior to said step of passing the fluid 17 through the passage means 3. The biological fluid sterilization method further comprising the step of introducing a protective agent. Method according to claim 13 or 14, characterized in that the residence time of the fluid (17) in the vessel (2) of the device (1) is 30 seconds to 10 minutes.