JP2003070904A - Filter holder and filter unit for separating organism component, and performance evaluation method of hemofiltration filter using the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a filter holder, and a filter unit for a organism component to be used for hemofiltration such as cellular segregation, leukocyte removal, and a performance evaluation method of a hemofiltration filter using the filter unit. SOLUTION: The filter holder is obtained by press-contacting a filter element housing member having a recess member for housing a filter element, a flange part and an opening part, and a lid member having a protrusion part to be fitted to the filter element housing member, a flange part and an opening part at the flange part of each member. Ring parts for inserting and fixing the filter element are provided respectively at the recess part of the housing member and the protrusion part of the lid member, and the protrusion part of the lid member is projected from the press-contacting surface of the flange toward the recess part side of the housing member. The filter element is housed in the filter holder to constitute the filter unit which never causes blood leakage and is good to handle. Further, the performance of the hemofiltration filter on the market can be evaluated by using the filter unit.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a filter holder for separating biological components used for blood filtration such as cell separation and leukocyte removal, a filter unit, and a method for evaluating the performance of a blood filtration filter using the filter unit.

[0002]

2. Description of the Related Art In recent years, in the field of blood transfusion, in order to prevent side effects after blood transfusion, it has become common to remove leukocytes from blood products by using a blood cell separation filter containing a non-woven fabric or a porous body. Is coming. It is strongly desired that the blood cell separation filter has high leukocyte removal performance and blood flowability and is stable.

In studying the performance design of such a desirable blood cell separation filter, normally, 200 to 500 ml of whole blood or blood products are passed through a prototype or an actual product to focus on the basic leukocyte removal performance. The performance evaluation is performed. For example, JP-A-2-13587 and JP-A-2-13588 disclose 400 whole blood containing CPD.
It is described that the evaluation was carried out using 100 ml or 500 ml, and it is described in JP-A No. 10-328295 and JP-A No. 11-328295.
Japanese Unexamined Patent Publication No. -20876 describes that evaluation is performed using 400 ml of blood. In both cases, actual products and prototypes of the same size were used for performance evaluation, and due to the filter size, a large amount of blood and blood products essential for medical treatment were required.

In the above evaluation, it is difficult to obtain a large amount of valuable blood products at one time, so it takes a considerable time to secure a sufficient evaluation score and obtain a highly accurate evaluation result. However, it has not been possible to satisfy the requirements for the comparative examination under the same desirable conditions for biological samples having large individual differences.

Therefore, an evaluation method using a small amount of blood has been tried, and in Japanese Patent Laid-Open Nos. 2000-51623 and 2000-197814, a filter element punched in a circular shape having a diameter of 25 mm is used as a dedicated filter. An evaluation method is described in which 5 ml of blood is passed through a column packed in a holder. Originally, in order to bring out the same performance as an actual blood filter, it is considered necessary to reduce the blood volume according to the cross-sectional area without changing the thickness of the filter in the filtration direction, but it is described in the above publication. None of the specifications and evaluation conditions of the filter holders taken into consideration considered faithful scale-down of performance evaluation. Therefore, these inventions are considered to be useful for initial screening to determine the tendency of the adsorptivity of platelets and the like between materials to some extent, but it is necessary to completely match the actual blood filter performance. Obviously, it cannot be used in performance design.

To solve the above problems, it is necessary to make the leukocyte removal filter itself small in size so that it can accurately cope with a small amount of blood or blood products so that the performance can be scaled down faithfully. . Moreover, at that time,
When blood passes through the filter, it cannot be accurately evaluated if there is blood leakage outside the effective cross-sectional area through which blood substantially passes. Therefore, a small filter unit that does not leak blood to the outside of the effective cross-sectional area has been demanded, and an appropriate filter holder has been required for that purpose.

In order to prevent liquid leakage outside the effective area,
Plastic filters and filter units in which the outer periphery of the filter is adhered and fusion-molded are already well known and widely used in various industries. However, a filter unit in which the outer periphery of the filter is adhered and welded is time-consuming and costly to manufacture, and when the used filter is taken out for analysis of blood flowability in the filter, adhesion between the housing and the filter, There is a drawback that the operation of disassembling the welded portion is complicated. Also,
There is a risk of accidentally damaging the filter when disassembling,
It was not suitable for use in order to efficiently advance the performance design of filters.

On the other hand, the filter and the housing are bonded together,
As a filter unit that does not weld, a type in which a filter is set in a screw type or pressure type filter holder is known, for example, a diameter of 13 mm.
Screw type filter holder (ADVANT, Japan
EC-made and screw type filter holder with a diameter of 25 mm (Japan, ADVANTEC-made, USA, CORNI
NG) or a pressure-holding type filter holder (made by ADVANTEC, Japan) having a diameter of 25 mm is commercially available. It is considered that blood performance can be evaluated by setting a blood filter using these filter holders and allowing a small amount of blood to pass through. In order to evaluate the blood filtration performance of a leukocyte removal filter that is actually used with a small amount of blood, the amount of non-woven fabric (thickness and number) for capturing leukocytes used in an actual leukocyte removal filter and the hemofiltration are used. It is conceivable to reduce the blood filtration cross-sectional area without changing the linear velocity.

However, as a result of examination by the present inventors, when a filter element having several tens or more non-woven fabrics laminated on these commercially available filter holders is set in the holder as in the case of the actual product leukocyte removal filter, the outer periphery of the filter is It was not possible to produce a product capable of filtering blood without causing any problems, such as side leakage or hemolysis. Commercially available filter holders are generally used for setting filter paper, and it is rather natural that a thick nonwoven fabric with tens of layers stacked could not be set reliably.

Therefore, in order to obtain the same performance as an actual blood removal filter, several non-woven fabrics which do not leak sideways even when blood is passed are set in the filter holder,
There is a method of connecting and using a plurality of filter holders so that the number of non-woven fabrics inserted in an actual leukocyte removal filter is the same. In this case, as the number of connected holders increases, the dead volume naturally increases, and the blood volume required for performance evaluation increases,
There is a problem that the flowability of blood changes and accurate blood filtration performance cannot be evaluated. If the number of non-woven fabrics that can be set per filter holder is large, the number of filter units to be connected can be reduced, so the problem of dead volume can be solved considerably, but such filter holders have not been known so far. .

That is, more than ten or more filter elements such as leukocyte removal filters can be set without welding and adhering the outer peripheral portion, and moreover, a filter which does not cause leakage of liquid containing biological components such as blood around the filter elements. The holder and the filter unit in which the filter element is inserted have heretofore been unknown. Further, no method has been known at all for evaluating the performance of a commercially available hemofiltration filter from the hemofiltration performance obtained by passing a small amount of blood through a filter unit.

[0012]

The object of the present invention is to solve the problems of the conventional blood evaluation, (1) without using a large amount of blood or blood products, and (2) dismantling the filter after use. We solved problems such as time and labor and filter damage,
(3) A filter holder that does not cause blood leakage around the filter element, and (4) is easy to handle.
And to provide a filter unit. Also,
It is intended to provide a method for evaluating the performance of a commercially available blood filtration filter by passing a small amount of blood through the filter unit.

[0013]

Means for Solving the Problems The inventors of the present invention have made extensive studies in order to solve the above problems. As a result, the filter element housing member having the recessed portion for housing the filter element, the flange portion, and the opening portion, and the lid member having the protruding portion, the flange portion, and the opening portion fitted into the recessed portion of the filter element housing member are mutually provided. In the filter holder that is pressed against the flange portion of, the ring portion for sandwiching and fixing the filter element is provided in each of the concave portion of the storage member and the convex portion of the lid member, and the convex portion of the lid member is the flange. By projecting from the pressure contact surface of to the concave side of the storage member,
The inventors have found that the above problems can be solved and completed the present invention. The present invention also relates to a biological component separating filter unit in which a filter element is sandwiched and fixed in such a filter holder. With the filter holder and the filter unit having this structure, a thick filter element, especially a laminated type filter element can be reliably set without liquid leakage.

Further, a filter element is set in this filter holder, and a filter unit having the same filtration thickness as a commercially available blood filtration filter is provided so that the blood volume per cross-sectional area and the blood filtration linear velocity are the same as the actual product. It was confirmed that when a small amount of blood was passed through, the same performance as that of a commercially available blood filtration filter was obtained with good reproducibility without causing blood leakage. The present invention also relates to a method for evaluating blood filtration performance using the biological component separating filter unit based on the above findings. In the filter holder and the filter unit of the present invention, the filter element can be securely sandwiched and fixed by the ring portion, and the filter cross-sectional area through which the liquid flows can be appropriately adjusted by selecting the inner diameter of the ring portion. It is particularly suitable for evaluating the performance of filters.

[0015]

DETAILED DESCRIPTION OF THE INVENTION The present invention will be described in detail below with reference to the drawings. FIG. 1 shows a front view of the filter holder of the present invention in a non-pressure contact state. The filter holder of the present invention is a combination of a pair of lid member 1 and filter element housing member 2. Each of the lid member 1 and the filter element storage member 2 has flange portions 6 and 6 each having a tapered outer peripheral portion.

The lid member 1 has a convex portion 4 which fits into a concave portion 5 which is a filter accommodating portion of the other filter element accommodating portion 2. The lid member 1 has an opening 7 so that a biological component such as blood can be introduced into the central portion of the filter element, and the ring portion 3 is provided concentrically with the opening 7. The other filter element housing member 2 has an opening 8 formed therein so that biological components such as blood can be led out from the central portion of the filter element, and the ring portion 3 is concentrically formed on the opening 8.
Is provided.

The ring portion 3 installed on the lid member 1 and the housing member 2 has a planar structure on the end faces 9 and 10 contacting the filter element, and when the lid member 1 and the housing member 2 are fitted together,
End faces 9 and 10 which contact the filter element of each other's ring part
Are designed to be an exact match. The lid member 1 and the filter element housing member 2 in which the filter element is housed in the concave portion 5 constitute a filter holder by combining the flanges 5 and 6 so as to face each other. Therefore, when the filter element is housed, the filter element has ring portions 3 and 3 installed on the lid member 1 and the housing member 2.
It will be fixed by being sandwiched between the end faces 9 and 10.

FIG. 2 shows an example of a sectional view of the filter holder shown in FIG. In the example shown in FIG. 2, the convex portion 4 of the lid member 1 and the concave portion 5 of the storage member 2 are provided with ring-shaped grooves 11 and 12 for mounting and fixing the ring 3.

Then, as shown in FIG. 3, by using the flange portions 6 and 6 of the filter holder and clamping them by the clamp 13, it is possible to make one filter unit having the filter element built therein.

A plurality of filter units are connected in series by inserting an opening 8 for leading out a biological component such as blood of one filter unit into an opening 7 for introducing a biological component such as blood of another filter unit. It is also possible to let.

The biological component of the present invention refers to blood, white blood cells, platelets, red blood cells, tissues, cells, proteins, plasma, nucleic acids, and bacterial cells. The biological component separating filter unit of the present invention can be applied to any solution as long as it is a solution containing the above. Considering the use of a filter element for filtering relatively large-sized ones such as cells, it can be more suitably used for blood, white blood cells, platelets, cells, and tissue-containing solutions. Furthermore, when considering the use of the filter element for leukocyte removal, it can be preferably used for blood, leukocyte, platelet, and erythrocyte-containing solution.

In the filter holder for separating biological components of the present invention, a pair of lid member and storage member are combined with each other at the flange portion and pressed together to store the filter element inside.
And when storing the filter element inside, in order to prevent liquid leakage from the filter element and fix it,
Ring portions need to be installed in the convex portions of the lid member and the concave portions of the housing member, and the convex portions need to protrude from the flange pressure contact surface.

For example, in the case of a thick non-woven fabric in which a dozen or more sheets are laminated like the filter element of a leukocyte removal filter, the filter element is set in the concave portion of the filter element storage member, and the lid member having the convex portion fitted into the concave portion is used. It is necessary to hold and pinch the filter element so that it does not shift. At this time, the filter element is substantially crimped, and
For the purpose of adjusting the portion where the liquid containing the biological component flows so as to be limited to a predetermined cross-sectional area, in the filter holder of the present invention, ring portions are provided in both the lid member convex portion and the storage member concave portion. In order to crimp and fix the filter element to the holder without a gap, it is necessary that the convex portion fitted into the concave portion protrudes from the flange pressure contact surface.

The ring portion installed on the filter holder may be integrally formed with the concave portion and the convex portion of the filter holder, or may be detachable. When conducting a filtration experiment using a small amount of blood, a smaller cross-sectional area is required.Therefore, the ring part is removable so that blood filtration performance can be evaluated according to the cross-sectional areas of various filter elements. More preferably, it is possible and replaceable.

More specifically, as the lid member molded body integrally forming the ring portion, the liquid is allowed to flow to the central portion of the disk-shaped molded body which coincides with the outer periphery of the recess for accommodating the filter element, In addition, a ring portion that protrudes in a convex shape from the surface of the disk-shaped molded body may be manufactured. Alternatively, the ring portion may be installed and fixed to the filter holder having one or more grooves for fixing the ring portion to the filter holder body. In addition, in order to confirm the installation of the ring part and prevent loss,
The ring portion is preferably colored so that it can be distinguished from the filter holder body.

Considering the ease of manufacturing the ring part and the handling property when setting the ring part and the filter element to the filter holder, the ring part has at least one groove which can be fixed to the filter holder body. It is more preferable that the ring portion is detachably fixed to the groove.

The inner diameter and outer diameter of the ring portion are not particularly limited, but in consideration of performing a small amount of blood filtration evaluation, the inner diameter is preferably 25 mm or less, more preferably 13 mm or less. Further, since the filter element is substantially clamped by the ring portion, the area where the ring portion faces the filter element is preferably small enough not to damage the filter element.
On the other hand, when the lid member and the storage member of the filter holder are put together, the ring portion installed on the lid member and the ring portion installed on the storage member are desired to be exactly the same, but in reality, molding process or Considering that a slight displacement occurs when the storage member and the lid member are aligned with each other, it is desirable that the difference between the outer diameter and the inner diameter of the ring portion has a certain width. . Taking the above into consideration, the difference between the outer diameter and the inner diameter of the ring portion is preferably in the range of 0.1 mm to 10 mm, more preferably in the range of 0.5 mm to 5 mm, and most preferably in the range of 1 to 2 mm.

Further, the ring portion provided on the lid member and the ring portion provided on the storage member have their end faces butted against each other to sandwich and fix the filter element. For that purpose, The end faces (9, 10) of the ring portion need to contact the filter element in a plane. The width of contact between the filter element and the end surface of the ring portion substantially corresponds to the difference between the outer diameter and the inner diameter of the ring portion, and the range of 0.1 mm to 10 mm is preferable, and the range of 0.5 mm to 5 mm. More preferred range,
The range of 1-2 mm is most preferable.

As the material of the molded body and the ring portion forming the filter holder, any material can be used as long as it can be molded such as plastic, metal, glass, ceramics and wood. Plastic is preferable in consideration of ease of molding and cost. Furthermore, considering that the fluidity of liquids containing biological components such as blood can be observed from the outside, relatively transparent materials are preferable, such as polymethylmethacrylate, polycarbonate, polypropylene, polyethylene, polystyrene, etc. More preferable.

The filter unit for separating biological components of the present invention can be used by pressing with the flange of the filter holder. As a method of pressure tightening, there are methods such as tightening bolts and nuts at several places on the outer periphery of the flange, a method using a clamp for sanitary piping connection, etc.
The method of clamping using a clamp is more preferable because it can be simply clamped.

The form of the filter element and the kind of material to be inserted into the filter unit of the present invention are not particularly limited, but it may be a porous body used for filtering a solution containing biological components such as blood and cells. Any form or material may be used.

Examples of the porous body include a non-woven fabric, a flat membrane, a hollow fiber, a woven fabric, a knitted fabric, a foamed body or a composite thereof. A non-woven fabric or a foam is generally used for the leukocyte removal filter, and the non-woven fabric or the foam is more preferable in consideration of performing filtration performance evaluation such as filtration of blood cells. Further, considering the ease of controlling the number of filter elements, a non-woven fabric is more preferable.

The material of the porous body is not particularly limited, but natural polymers such as cotton, hemp and silk, nylon, polyester, polyacrylonitrile, polyolefin, halogenated polyolefin, polyurethane, polysulfone,
Polyether sulfone, poly (methacrylate), ethylene-polyvinyl alcohol copolymer, butadiene-
A synthetic polymer such as an acrylonitrile copolymer or a mixture thereof may be considered.

The thickness of the filter element that can be inserted into the biological component separating filter holder of the present invention is not particularly limited, but when the filter element is set in the filter holder concave portion and the other lid member is pressure-contacted, it is clamped. Any thickness is possible. In addition, there should be no leakage at the periphery of the filter element when the solution, such as blood, is passed under pressure. As a result of investigating the relationship between the filter thickness when pressed and blood leakage, the present inventors have found that the filter element inserted into the filter unit for treating a biological component of the present invention has a pressed thickness of 0.05. It has been found that a range of ~ 2 mm is preferred. More preferably,
The range is 0.5 mm to 1.5 mm.

As a method for injecting blood into the filter unit of the present invention, there are a drop method and a constant flow method. The head difference referred to here is the distance between the blood bag and the collection bag in the filtration system in which the blood cell separation filter is connected by a blood circuit or the like between the blood bag and the collection bag. Such filtration methods are common in blood tests,
Although it is a preferable method also in the filter unit of the present invention, a constant drop condition is set because the drop causes a change in the flow rate into the blood cell separation filter and further tends to affect the blood cell separation filter. It is preferable.

As a method using a constant flow rate, there is a method in which a device such as a pump is installed on the inlet side of the blood cell separation filter to allow it to flow into the blood cell separation filter. In this case, since the blood flows into the blood cell separation filter at a low flow rate, there is no change in the processing time. Therefore, it is preferable to measure the pressure loss of the blood cell separation filter. For example, there is a filtration method in which a pressure monitor is provided at the inlet and outlet of the blood cell separation filter to flow blood at a constant flow rate, and the pressure loss is measured when the pressure at the inlet of the blood cell separation filter is stable. It is not limited to the examples. The filtration linear velocity of both methods is preferably 10 cm / min or less, more preferably 1 cm / min or less.

When blood is filtered using the filter holder of the present invention, blood may be introduced from the opening on the side of the lid member having the convex portion and the blood may be recovered from the opening on the side of the filter element storage member, or vice versa. Alternatively, blood may be flown with the filter element housing member side upstream and the lid member side downstream.

[0038]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto.

Example 1 (Preparation of filter holder and filter unit) A filter holder made of polymethylmethacrylate was prepared according to FIG. The lid member 1 and the filter element accommodating member 2 had an outer diameter of 50 mm and a diameter of a concave portion and a convex portion of 20 mm. The ring portions 3 and 3 installed in the filter holder were made of polymethylmethacrylate material having inner diameters of 7 mm, 10 mm, and 13 mm and outer diameters of 11 mm, 14 mm, and 17 mm, respectively.

[0039]

[Example 2 and Comparative Example 1] (Nonwoven fabric insertion test) Example 1
Asahi Medical's leukocyte removal filter "Sepacell" punched into a circular shape with a diameter of 20 mm using a cutting machine "SDL-100" (manufactured by Dumbbell, Japan) in a filter holder with an inner diameter of the ring portion of 13 mm manufactured in 1. ^TM RZ-
The main constituent non-woven fabrics used in "2000" are 16, 2
It was tested whether or not it could be set by stacking 0 and 30 sheets and clamping them with a clamp. As a comparative example, a commercially available filter holder "KS-25F" of a type that is clamped with a clamp.
The same test was performed on (manufactured by ADVANTEC, Japan). The results are shown in Table 1. The non-woven fabric was able to be stacked and set up to 30 pieces on the filter holder of Example 1, but as a comparative example, the commercially available filter holder "KS-25F" (manufactured by ADVANTEC, Japan) had only 20 pieces. I couldn't set it.

[0040]

[Table 1] Non-woven fabric insertion test ──────────────────────────────────── Number of filters Example 2 Comparative example 1                   Holder made in Example 1 Commercially available holder "KS-25F" ────────────────────────────────────   16 sheets filter set possible filter set possible   20 sheets filter set possible filter set possible   30 pieces filter set possible filter set not possible ────────────────────────────────────

Further, the thickness of the filter element was measured when the non-woven fabric prepared in Example 1 was inserted into the filter holder in an overlapped manner and clamped with a clamp. The results are shown in Table 2. The maximum thickness of 30 sheets was 1.65 mm.

[Table 2] Thickness of the filter element when the filter was inserted into the filter holder prepared in Example 1 and clamped with a clamp ─────────────────────── ─────────────── Example 2 Number of filters Thickness of filter element when pressed (mm) ─────────────────── ────────────────── 8 sheets 0.43 16 sheets 0.88 20 sheets 1.09 30 sheets 1.65 ──────────── ────────────────────────

[0042]

Example 3 and Comparative Example 2 (Blood Leakage Test) 150 ml of blood was collected from a healthy volunteer using a 19G winged needle, immediately transferred to a blood bag containing 21 ml of the anticoagulant CPD solution, and mixed well. Blood bag is COOL I
NCUBATOR "PCl-300" (manufactured by Inai Seieido Co., Ltd., Japan) was stored at 20 [deg.] C. Using the cutting machine "SDL-100" (manufactured by Dumbbell, Japan) in the filter holder of Example 1, a white blood cell removal filter "Sepacell ^™ " manufactured by Asahi Medical Co., which was punched into a circle with a diameter of 20 mm.
The main constituent non-woven fabric used in RZ-2000 "is 1
6 sheets were stacked and set. In addition, as a comparative example, a commercially available filter holder “KS-25F” (ADVA, Japan)
16 sheets of non-woven fabric punched into a disc shape having a diameter of 20 mm were similarly stacked and set on the NTEC company).

After 3 hours, it was confirmed that the blood temperature was 20 ° C., and 15.5 ml was added to each 20 ml syringe for each test section.
Blood was collected. Two connected filter units,
Syringe pump "55-2219" with the outlet facing up
(Manufactured by HARVARD, USA) and a syringe containing blood was connected. The filtration linear velocity is 0.56 cm / m
The flow rate was set to 0.739 ml / min so as to be in, the blood was flown into the filter unit by pushing out with a pump, and when the blood flowed, 13.3 ml was collected, the operation was stopped.

The results are shown in Table 3. It was confirmed that the filter unit in which the nonwoven fabric was set in the filter holder of Example 1 had sufficient blood flow and no blood leakage. On the other hand, a commercially available filter holder "KS-25"
F ”(manufactured by ADVANTEC, Japan) with 16 non-woven fabrics had blood leakage and hemolysis. In addition, it was confirmed that, in the case where 20 sheets of non-woven fabric were inserted, the blood stopped flowing 7 and 8 minutes after the blood started to flow, and hemolysis occurred.

[0045]

[Table 3] Blood leak test ───────────────────────────────────   Filter unit Number of non-woven fabrics Results of hemofiltration test ─────────────────────────────────── Example 3   16 filter units of Example 1 No blood leakage                                 20 sheets No blood leak                                 30 sheets There is a slight blood leak (bleeding) Comparative example 2   16 commercially available filter units with blood leak and hemolysis     "KS-25F" 20 sheets Blood stops flowing in 7 or 8 minutes                                           It was There is hemolysis. ───────────────────────────────────

[0046]

[Example 4 and Comparative Example 3 and Reference Example 1] (Evaluation test of blood filtration performance) A whole blood product having 16 main sheets, 32 pieces of which are composed of two filter units which are confirmed to have no blood leakage. White blood cell depletion filter product "Sepacel ^TM RZ-200" that can process 2 units (450 ml)
It was examined whether the hemofiltration performance evaluation corresponding to "0" was possible. The effective cross-sectional area of the product "Sepaseru ^TM RZ-2000" is 45cm ^2, the effective cross-sectional area in the filter unit of this embodiment is for a 1.33 cm ^2, collected blood volume reduction 1 33.8 min It was carried out in. 150 ml of blood was collected from a healthy volunteer using a 19G winged needle, immediately transferred to a blood bag containing 21 ml of the anticoagulant CPD solution, and mixed well. Blood bag is COLOR IN
CUBATOR "PCl-300" (manufactured by Inai Seieido Co., Ltd., Japan) was stored at 20 ° C.

A cutting machine "SDL-100" (manufactured by Dumbbell, Japan) was used for the filter unit of Example 1.
16 pieces of the main constituent non-woven fabrics used in the Asahi Medical's leukocyte removal filter "Sepacell ^TM RZ-2000" punched into a disc shape with a diameter of 20 mm are set, and this filter unit is connected in two, the main constituent A filter unit having a non-woven fabric of 32 sheets, which is the same as that of the product "Sepacell ^™ RZ-2000", was prepared. 24 hours later,
It was confirmed that the blood was at 20 ° C., and 15.5 ml of blood was collected in a 20 ml syringe. Place the two connected filter units with the outlet facing up and the syringe pump “55
-2219 "(manufactured by HARVARD, USA), and a syringe containing blood was connected. The filtration linear velocity is 0.
The flow rate is 0.739 ml / so that it becomes 56 cm / min.
It was set to ml, pushed out by a pump, blood was poured into the filter unit, and the operation was stopped when 13.3 ml was collected. When hemofiltration was performed using the filter unit of the present example, and 13.3 ml of the collected filtered blood was subjected to a blood experiment with an actual product-level leukocyte removal filter using 2 units (450 ml of whole blood). Equivalent to the recovery amount of.

A part of the blood after filter filtration is transferred to an Eppendorf tube, and a blood cell measuring device "MAXM A
/ LRetic "(manufactured by Beckman Coulter, Inc., USA)
It was measured at. In addition, since the minute white blood cell count is below the detection limit of the measuring device, "LeucoCOUN"
"T ^TM " kit (manufactured by Becton Dickinson, USA)
Flow cytometer "FACS Ca
The number of white blood cells stained with nucleic acid was counted when 30,000 internal standard fluorescent beads were counted by "libur". The leukocyte-removing ability and the platelet recovery rate were calculated by the following equations 1 and 2.

[0049]

[Equation 1]

[0050]

[Equation 2]

Table 4 shows the results of the leukocyte-removing ability, the erythrocyte recovery rate, and the platelet recovery rate after filtration after 24 hours. As a reference example, one different from the present embodiment
Using 450 ml of blood preserved daily, and using the product of the leukocyte removal filter, "Sepacell ^™ RZ-2000",
Table 4 also shows the results of leukocyte-removing ability, erythrocyte recovery rate, and platelet recovery rate when filtered by cm. Further, as a comparative example, four non-woven fabrics used in the examples were set in a commercially available screw type filter holder having a diameter of 25 mm (manufactured by CORNING, USA), and blood was drawn at a linear velocity of 0.56 cm / mi.
Table 4 also shows the results of leukocyte depletion ability, erythrocyte count rate, and platelet recovery rate when the amount of blood collected was 6 ml.
It was shown to.

[0052]

[Table 4] Hemofiltration evaluation test ────────────────────────────────── Measurement item Example 4 Reference Example 1 Comparative Example 3 ────────────────────────────────── Leukocyte removal ability [−log] 4.32 4.29 1.77 Red blood cell recovery rate [%] 101 102 100 Platelet recovery rate [%] 1.4 2.8 40.4 ──────────────────────── ─────────── From the results in Table 4, the leukocyte-removing capacity and platelet recovery rate evaluated by hemofiltration with the filter unit of the Example are very good as the results obtained using the actual product. It was confirmed that they match. On the other hand, although four non-woven fabrics were set in a commercially available filter holder, the leukocyte-removing ability and the platelet recovery rate did not match at all.

[0053]

Example 5 and Reference Example 2 (Hemofiltration test) 150 ml of blood was collected from two healthy volunteers using an 18G winged needle, immediately transferred to a blood bag containing 21 ml of the anticoagulant CPD solution, and mixed well. . Furthermore, aseptically 40
Blood for 2 persons was transferred to a blood bag for collecting 0 ml and mixed well, and then COOL INCUBATOR "PCl-30"
0 "(manufactured by Inai Seieido Co., Ltd., Japan) at 20 ° C.

As a reference example, a product "Sepacel ^™ RZ-100" having 26 main constituent nonwoven fabrics for treating 1 unit of blood is used.
0 ”was used. After 3 hours, it was confirmed that the blood was at 20 ° C., 1 unit of 228 ml was transferred to the bag on the upper supply side, and filtration was started when the difference from the collection bag was 90 cm. The filtration is completed in 13 minutes and 1 second, and the average processing speed is 1.
It was 6.1 ml / min.

As an example corresponding to this, with the same product
There is "Separcel^TMRZ-1000 ”container was disassembled and
Take out the woven cloth, and use the cutting machine "SDL-100" (Japan,
It was punched into a circular shape with a diameter of 20 mm with a dumbbell company.
This non-woven fabric has 26 filters to 13 filters and 2 filters each.
Separately set in units and connected in two, with the main structure non-woven
The number of cloths is the product "Sepacel^TMSame structure as RZ-1000 "
I made it Product "Separcel^TMRZ-1000 "blood
Immediately after completion of the liquid filtration, 20 ml of syrin
11 ml of blood was collected in the test tube and two filter units were connected.
The syringe pump "55-221
9 "(Harvard, USA)
The syringe containing was connected. Product "Separcel^TMRZ-
To match the blood filtration linear velocity with 1000 ”,
Set to 0.63 ml / ml, push out with a pump,
Blood was poured into the Luther unit, and 8.6 ml was collected.
At that point, the operation was stopped. The recovery volume of 8.6 ml is
Product "Separcel ^TMRZ-1000 "effective area and fill
24. Calculated from the ratio of the effective cross-sectional areas in the
It is a value reduced to 1/4. Blood after filtration
Was analyzed in the same manner as in Example 4, and the leukocyte-removing ability,
Platelet recovery rate and red blood cell recovery rate were calculated.

The results are shown in Table 5. As a result, the value obtained by performing hemofiltration evaluation using the product "Sepacel ^™ RZ-1000" and the value obtained using the filter holder made corresponding to this were in very good agreement.

Table 5: Filter unit of the present invention and RZ-10
Hemofiltration Test Using 00 ───────────────────────────────── Measurement Item Example 5 Reference Example 2 ── ─────────────────────────────── Leukocyte removal capacity [−log] 4.99 4.49 Red blood cell recovery rate [%] 99.3 102.5 Platelet recovery rate [%] 0.0 0.0 ───────────────────────────────── ─

[0057]

The biological component separating filter unit of the present invention does not cause blood leakage around the filter element,
Moreover, it is a filter unit that can be easily disassembled without damaging the filter, and the performance of the actual blood separation filter can be accurately evaluated with a small amount of blood.

[Brief description of drawings]

[Figure 1] Front view of the filter holder

FIG. 2 is a sectional view of the filter holder.

FIG. 3 is an external view of a filter unit.

[Explanation of symbols]

1 Lid member 2 Filter element storage member 3 Ring part 4 convex 5 recess 7,8 openings 9,10 Ring end face 11, 12 Groove for installing ring part 13 clamps

Continued front page    F-term (reference) 4C077 AA12 BB02 CC05 EE01 KK11                       LL02 NN02 PP01 PP02 PP07                       PP08                 4D019 AA03 BA12 BA13 BB02 BB03                       BB07 BD01 CB01 CB02                 4D064 AA29 BQ01

Claims (12)

[Claims] 1. A filter element accommodating member having a concave portion for accommodating a filter element, a flange portion and an opening portion, and a lid member having a convex portion, a flange portion and an opening portion which fit into the concave portion of the filter element accommodating member. In the filter holder in which the flange portions are pressed against each other, a ring portion for sandwiching and fixing the filter element is installed in each of the concave portion of the storage member and the convex portion of the lid member, and the lid member The filter holder for separating biological components, characterized in that the convex portion of the convex portion projects from the pressure contact surface of the flange toward the concave portion of the storage member. 2. The filter holder for separating biological components according to claim 1, wherein the ring portion and the filter element are in contact with each other on a flat surface because the end surface of the ring portion has a flat surface. 3. The filter holder for separating biological components according to claim 2, wherein the contact plane between the ring portion and the filter element has a width of 0.1 to 10 mm. 4. The ring portion is attachable and detachable, and the concave portion of the filter element housing member and the convex portion of the lid member have at least one groove for mounting the ring portion on the filter holder. The filter holder for biological component separation according to any one of claims 1 to 3, characterized in that. 5. The filter holder for separating biological components according to claim 1, wherein the filter element housing member and the lid member are made of plastic. 6. The filter holder for separating biological components according to claim 1, wherein the ring portion is made of plastic. 7. The filter holder for separating biological components according to claim 1, wherein the inner diameter of the ring portion is 25 mm or less. 8. A filter unit for separating biological components, wherein a filter element is sandwiched and fixed to the filter holder for separating biological components according to any one of claims 1 to 7. 9. The filter unit for separating biological components according to claim 7, wherein the flange portion of the filter holder is clamped with a clamp. 10. The filter unit for separating biological components according to claim 8, wherein the filter element is a non-woven fabric. 11. The biological component separation filter unit according to claim 8, wherein the thickness of the filter element when sandwiched and fixed is 0.05 mm or more and 2 mm or less. 12. A method for evaluating the performance of a blood filtration filter, which comprises using the filter unit for separating biological components according to any one of claims 8 to 11.