Plasma filtering composite material applied in combination with wheat germ agglutinin and plasma separating device

Abstract

The invention provides a plasma filtering composite material applied in combination with wheat germ agglutinin and a non-centrifugal plasma separation device, and belongs to the technical field of medical apparatuses and instruments. The invention provides a plasma filtering composite material applied in combination with wheat germ agglutinin, which is of a multi-layer structure and sequentially comprises a filter screen with the pore diameter of 38-42 microns, a first filter membrane with the pore diameter of 18-22 microns, a second filter membrane with the pore diameter of 8-12 microns and a third filter membrane with the pore diameter of 0.8-1.2 microns from top to bottom. Wheat germ agglutinin (WGA) can be combined with a specific receptor on the surface of red blood cells (RBC) to promote rapid agglutination reaction of the RBC. By reasonably designing the structures of the filter screen and the filter membrane of the plasma filtering composite material, red blood cells agglutinated by WGA and large-particle components in blood can be effectively intercepted, and the plasma filtering efficiency is improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical devices, and in particular relates to a plasma filtration composite material and a plasma separation device combined with wheat germ agglutinin. Background Art

[0002] Traditional plasma separation methods primarily rely on centrifugation, which separates blood cells from plasma through centrifugal force. However, centrifugation is equipment-intensive, complex, and time-consuming, limiting its applicability in resource-limited or emergency medical settings. Therefore, developing efficient, simple, and centrifuge-free plasma separation methods has become an important research direction.

[0003] Currently, the primary non-centrifugal plasma separation method is membrane plasma separation. Membrane plasma separation utilizes a semipermeable membrane with a specific pore size to separate plasma. As blood flows through the membrane, plasma can pass through and be separated, while blood cells and other components cannot. However, the filtration efficiency of existing membrane plasma separation methods needs to be further improved. Summary of the Invention

[0004] The object of the present invention is to provide a plasma filtration composite material and a plasma separation device combined with wheat germ agglutinin. The solution of the present invention can improve the filtration efficiency of membrane plasma separation.

[0005] The present invention provides a plasma filtration composite material for combined use with wheat germ agglutinin. The plasma filtration composite material has a multilayer structure, and comprises, from top to bottom, a filter screen, a first filter membrane, a second filter membrane, and a third filter membrane; the pore size of the filter screen is 38-42 μm; the pore size of the first filter membrane is 18-22 μm; the pore size of the second filter membrane is 8-12 μm; and the pore size of the third filter membrane is 0.8-1.2 μm.

[0006] Preferably, the materials of the first filter membrane, the second filter membrane and the third filter membrane include glass cellulose membrane.

[0007] The present invention also provides a plasma filter comprising a filter cavity; the plasma filtration composite material described in the above solution is installed in the filter cavity.

[0008] The present invention also provides a plasma separation device, comprising the plasma filter and plasma collector described in the above scheme; a bleeding port is provided at the lower portion of the filter cavity; and the plasma collector is connected to the bleeding port through a pipeline.

[0009] Preferably, the plasma collector comprises a suction type plasma collector; the suction direction of the plasma collector is from bottom to top; the plasma collector comprises a syringe.

[0010] The present invention also provides use of the plasma filtration composite material, the plasma filter or the plasma separation device described in the above scheme in plasma separation.

[0011] The present invention also provides the use of wheat germ agglutinin in combination with the plasma filtration composite material, the plasma filter or the plasma separation device described in the above scheme in plasma separation.

[0012] Preferably, the wheat germ agglutinin is used at a concentration of 0.36-0.45 g / L.

[0013] The present invention also provides a method for separating plasma, comprising the following steps: Blood is mixed with wheat germ agglutinin and agglutinated; Filtering the blood after the coagulation treatment using the plasma filtration composite material, the plasma filter or the plasma separation device described in the above scheme; The filtered fractions were collected to obtain plasma.

[0014] Preferably, the wheat germ agglutinin is used at a concentration of 0.36-0.45 g / L.

[0015] The present invention provides a plasma filtration composite material that utilizes wheat germ agglutinin. The plasma filtration composite material has a multilayer structure, comprising, from top to bottom, a filter screen, a first filter membrane, a second filter membrane, and a third filter membrane. The filter screen has a pore size of 38-42 μm; the first filter membrane has a pore size of 18-22 μm; the second filter membrane has a pore size of 8-12 μm; and the third filter membrane has a pore size of 0.8-1.2 μm. Wheat germ agglutinin (WGA) binds to specific receptors on the surface of red blood cells (RBCs), inducing rapid agglutination of RBCs. The resulting RBC aggregates are significantly larger than individual RBCs. By rationally designing the filter screen and filter membrane structure of the plasma filtration composite material, the present invention effectively intercepts WGA-agglutinated RBCs and large particles in the blood, thereby improving plasma filtration efficiency. Therefore, the synergistic effect of the plasma filtration composite material and WGA can achieve plasma separation and improve separation efficiency. It has been verified that the plasma filtration composite material can be used to filter the lectin-treated blood, and the initial separation of plasma and cells can be completed after standing for 30 seconds. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0017] Figure 1 Schematic diagram of the structure of the plasma separation device of the present invention; Figure 2 The agglutination of RBCs under different WGA concentrations; Figure 3 This is the plasma separation flow chart; Figure 4 This is the principle diagram of plasma separation; Figure 5 The fibrinogen concentration in the plasma obtained by centrifugation and filtration was tested by an automated coagulation analyzer, and then analyzed by a paired t-test. The blue color represents the result before filtration, the red color represents the result after filtration, and the orange-yellow color represents the difference between the two. DETAILED DESCRIPTION

[0018] The present invention provides a plasma filtration composite material for combined use with wheat germ agglutinin. The plasma filtration composite material has a multilayer structure, and comprises, from top to bottom, a filter screen, a first filter membrane, a second filter membrane, and a third filter membrane; the pore size of the filter screen is 38-42 μm; the pore size of the first filter membrane is 18-22 μm; the pore size of the second filter membrane is 8-12 μm; and the pore size of the third filter membrane is 0.8-1.2 μm.

[0019] As an embodiment, the plasma filtration composite material consists of a filter screen, a first filter membrane, a second filter membrane and a third filter membrane from top to bottom.

[0020] As an embodiment, the pore size of the filter screen is 40 μm; the pore size of the first filter membrane is 20 μm; the pore size of the second filter membrane is 10 μm; and the pore size of the third filter membrane is 1 μm.

[0021] Wheat germ agglutinin (WGA) binds to specific receptors on the surface of red blood cells (RBCs), prompting rapid RBC agglutination, thereby improving filtration efficiency. The plasma filtration composite material of the present invention utilizes a filter screen and gradient filter membrane to form a composite. The multi-layered structure utilizes the physical sieving effect to gradually intercept large particles in the blood, including RBCs and RBC aggregates, white blood cells, and platelets. Therefore, through the rational design of the filter screen and membrane structure of the plasma filtration composite material, the present invention effectively intercepts RBCs and large particles in the blood that have been agglutinated by WGA, thereby improving plasma filtration efficiency.

[0022] As an embodiment, the filter screen includes a nylon filter screen.

[0023] As an embodiment, the material of the first filter membrane, the second filter membrane and the third filter membrane includes glass fiber membrane. The glass fiber membrane has a weak adsorption capacity for proteins in plasma, which is conducive to the retention of plasma protein components.

[0024] The present invention also provides a plasma filter comprising a filter cavity; the plasma filtration composite material described in the above solution is installed in the filter cavity.

[0025] As an embodiment, a blood inlet is provided at the upper portion of the filter cavity. Furthermore, the upper portion of the filter cavity is open.

[0026] As an embodiment, the plasma filter further includes a supporting mesh plate, the mesh of which is used for plasma filtration; the plasma filtration composite material is supported on the supporting mesh plate and both are accommodated in the filtration cavity; the edges of the plasma filtration composite material are fixed by gluing to ensure its airtightness.

[0027] The present invention also provides a plasma separation device, comprising the plasma filter (1) and the plasma collector (2) described in the above scheme; a bleeding port is provided at the lower portion of the filter cavity; the plasma collector is connected to the bleeding port via a pipeline; the structural diagram of the plasma separation device is shown in FIG. Figure 1 shown.

[0028] As an embodiment, the plasma collector includes a suction type plasma collector; the suction direction of the suction type plasma collector is from bottom to top; the suction type plasma collector includes a syringe.

[0029] The plasma filter of the present invention, combined with the aspiration operation of the aspiration-type plasma collector, can rapidly obtain high-purity plasma. Furthermore, the aspiration-type plasma collector utilizes negative pressure to draw plasma, effectively obtaining plasma while also reducing the hemolysis rate. This has no significant impact on FIB concentration testing, and can, to a certain extent, meet the clinical needs of plasma.

[0030] The present invention also provides the use of the plasma filtration composite material, the plasma filter or the plasma separation device described in the above scheme in plasma separation.

[0031] The present invention also provides the use of wheat germ agglutinin in combination with the plasma filtration composite material, the plasma filter or the plasma separation device described in the above scheme in plasma separation.

[0032] As an embodiment, the plasma separation is plasma separation of a whole blood sample.

[0033] As an embodiment, the wheat germ agglutinin is used at a concentration of 0.36-0.45 g / L.

[0034] The above scheme is a method for plasma separation, comprising the following steps: Blood is mixed with wheat germ agglutinin and agglutinated; Filtering the blood after the coagulation treatment using the plasma filtration composite material, the plasma filter or the plasma separation device described in the above scheme; The filtered fractions were collected to obtain plasma.

[0035] As an embodiment, the wheat germ agglutinin is used at a concentration of 0.36-0.45 g / L.

[0036] The technical solution of this invention not only simplifies the plasma separation process but also shortens processing time and reduces equipment dependency, providing a new solution for point-of-care testing (POCT), remote healthcare, and immediate diagnosis. Further optimizing the synergistic effect of the membrane material and biological reagents based on this technical solution is expected to expand its application potential in clinical blood transfusion, disease screening, and other fields.

[0037] To further illustrate the present invention, a plasma filtration composite material combined with wheat germ agglutinin and a non-centrifugal plasma separation device provided by the present invention are described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0038] Test Example 1 1. Test materials and process WGA was accurately weighed using an electronic analytical balance and dissolved in physiological saline to prepare final concentrations of 1 g / L, 2 g / L, 3 g / L, 4 g / L, and 5 g / L. Blood samples were collected using vacuum tubes containing 1:9 sodium citrate. After thorough mixing, 200 μL of whole blood was accurately pipetted into EP tubes. Six groups were included in the experiment. The control group received 20 μL of physiological saline, while the remaining five experimental groups received 20 μL of the aforementioned WGA solutions of varying concentrations (1 to 5 g / L). The mixed blood samples were quickly transferred to blood typing cards, and the agglutination of each WGA solution concentration was recorded using a Deli high-speed scanner.

[0039] 2. Test results The test results can be found in Figure 2 .

[0040] Adding wheat germ agglutinin (WGA) to the blood can promote red blood cell (RBC) aggregation. The results showed that when the WGA concentration in the blood was 0.09 g / L, RBC agglutination was weakly positive; as the WGA concentration in the blood increased, the degree of RBC agglutination gradually increased; when the WGA concentration in the blood reached 0.36 g / L, its agglutination score reached "++++", indicating that the RBC agglutination effect was optimal.

[0041] Experimental Example 2: Verify whether WGA will cause RBC hemolysis and thus affect the accuracy of the test results 1. Test materials and process The hemolysis rate of RBC in WGA solution of different concentrations was measured to evaluate it. The steps are as follows: WGA was accurately weighed using an electronic analytical balance and dissolved in physiological saline to prepare final concentrations of 1 g / L, 2 g / L, 3 g / L, 4 g / L, and 5 g / L. Blood samples were then collected using EDTA-K2 vacuum tubes and gently mixed. Hemoglobin (Hb) content and HCT were measured using an automated hematology analyzer. Subsequently, 200 μL of whole blood was accurately pipetted into 1.5 mL EP tubes. Six groups were included in the experiment. The control group received 20 μL of physiological saline, while the remaining five experimental groups received 20 μL of the WGA solutions of varying concentrations (1 to 5 g / L). After gentle mixing, the samples were centrifuged at 2000 rpm for 5 min in a small high-speed refrigerated centrifuge. After centrifugation, 270 μL of Tris-HCl buffer (pH = 8.0) was added to a 96-well plate, followed by 30 μL of the supernatant and thorough mixing. The absorbance (OD value) was measured at 380 nm, 415 nm, and 450 nm using a multifunctional microplate reader. The RBC hemolysis rate was calculated using the following formula: 2. Test results The results are shown in Table 1.

[0042] Table 1 Effects of different WGA concentrations on RBC hemolysis rate

[0043] As WGA concentration in the blood increased, the RBC hemolysis rate showed only a slight increase. At a WGA concentration of 0.36 g / L, the hemolysis rate reached a maximum of 0.30%. Clinical transfusion standards require a hemolysis rate below 0.8%, while laboratory testing requires a hemolysis rate below 2%. Therefore, the degree of RBC hemolysis caused by 0.36 g / L WGA in the blood was relatively mild and fully met the experimental requirements.

[0044] Example 1 Whole blood sample plasma separation Plasma separation device: composed of a plasma filter and a syringe; the plasma filter consists of a filter chamber, a supporting mesh plate and a plasma filtration composite material; a blood inlet is provided at the top of the filter chamber; the plasma filtration composite material is supported on the supporting mesh plate and both are accommodated in the filter chamber; the edges of the plasma filtration composite material are fixed by glue to ensure its airtightness; a bleeding port is provided at the bottom of the filter chamber; the syringe is connected to the bleeding port through a pipeline, and the syringe draws suction from bottom to top; the plasma filtration composite material consists of a filter screen (made of nylon), a first filter membrane (made of glass cellulose membrane), a second filter membrane (made of glass cellulose membrane) and a third filter membrane (made of glass cellulose membrane) from top to bottom; the pore size of the filter screen is 40 μm; the pore size of the first filter membrane is 20 μm; the pore size of the second filter membrane is 10 μm; and the pore size of the third filter membrane is 1 μm.

[0045] Whole blood sample: from Beijing Friendship Hospital, the volume is about 200 μL.

[0046] First, 20 μL of 4 g / L WGA solution was added to 200 μL of whole blood sample and gently mixed, and then added to the plasma separation device.

[0047] This embodiment utilizes the property of WGA binding to specific receptors on the surface of RBCs to induce a rapid agglutination reaction of RBCs. When an appropriate amount of WGA is added to a blood sample, WGA can cause RBCs to rapidly form aggregates. The size of these RBC aggregates will be significantly larger than that of a single RBC. They are filtered through a plasma filter, and then the plasma is aspirated from the bleeding site using a syringe. This allows the RBC aggregates to be efficiently retained when passing through each layer of the filter membrane. This process can improve the removal efficiency of RBCs in the blood, ensure the rapid separation of plasma components, and further expand the application range of this POCT detection method. For a flow chart of plasma separation, see Figure 3 , plasma separation principle diagram see Figure 4 .

[0048] Test Example 3 1. Test materials and process WGA was accurately weighed using an electronic analytical balance and dissolved in physiological saline to prepare a WGA solution with a final concentration of 4 g / L. Subsequently, blood samples were collected using EDTA-K2 vacuum tubes. After gentle mixing, the hemoglobin (Hb) content and HCT of the blood samples were determined using an automated hematology analyzer. Subsequently, 200 μL of whole blood was accurately pipetted into 1.5 mL EP tubes, and 20 μL of the 4 g / L WGA solution was added to achieve a final WGA concentration of 0.36 g / L. After gentle mixing, the blood was filtered through a filtration device. After obtaining filtered plasma, 270 μL of Tris-HCl buffer (pH = 8.0) was added to a 96-well plate, followed by 30 μL of filtered plasma and thorough mixing. The absorbance (OD value) was measured at wavelengths of 380 nm, 415 nm, and 450 nm using a multifunctional microplate reader. The RBC hemolysis rate was calculated according to the following formula: 2. Test results The test results are shown in Table 2.

[0049] Table 2 RBC hemolysis rate results after filtration of blood containing 0.36 g / L WGA

[0050] The results showed that the average hemolysis rate caused by the plasma prepared in Example 1 was 0.61%, indicating mild hemolysis.

[0051] Test Example 4 1. Test materials and process Fifteen clinical blood samples were analyzed. The plasma obtained by centrifugation before filtration and the plasma obtained after filtration were tested using an automatic coagulation analyzer to observe the effect on the fibrinogen (FIB) concentration.

[0052] The whole blood sample was divided into two parts, one of which was separated into plasma by centrifuge and the FIB concentration result was detected by a fully automatic coagulation analyzer. The other blood sample was then filtered through the device, and the filtered plasma was detected by a fully automatic coagulation analyzer. The changes in the FIB detection concentration of the plasma before and after filtration were compared.

[0053] 2. Test results The paired t-test results are as follows Figure 5 As shown, there was no significant effect of plasma before and after filtration on FIB concentration ( p >0.05), indicating that Example 1 achieved effective filtration of plasma.

[0054] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. A plasma filtration composite material combined with wheat germ agglutinin, characterized in that: The plasma filtration composite material is a multi-layer structure, which comprises, from top to bottom, a filter screen, a first filter membrane, a second filter membrane and a third filter membrane; The pore size of the filter is 38-42 μm; The pore size of the first filter membrane is 18-22 μm; The pore size of the second filter membrane is 8-12 μm; The pore size of the third filter membrane is 0.8-1.2 μm.

2. The plasma filtration composite material according to claim 1, characterized in that The materials of the first filter membrane, the second filter membrane and the third filter membrane include glass cellulose membrane.

3. A plasma filter, characterized in that It comprises a filtration cavity; the plasma filtration composite material according to claim 1 or 2 is installed in the filtration cavity.

4. A plasma separation device, characterized in that: It comprises the plasma filter and plasma collector according to claim 3; a bleeding port is provided at the lower portion of the filter cavity; and the plasma collector is connected to the bleeding port through a pipeline.

5. The plasma separation device according to claim 4, characterized in that: The plasma collector comprises a suction type plasma collector; the suction direction of the plasma collector is from bottom to top; the plasma collector comprises a syringe.

6. Use of the plasma filtration composite material according to claim 1 or 2, the plasma filter according to claim 1 or 2, or the plasma separation device according to claim 4 or 5 in plasma separation.

7. Use of wheat germ agglutinin in combination with the plasma filtration composite material according to claim 1 or 2, the plasma filter according to claim 3, or the plasma separation device according to claim 4 or 5 in plasma separation.

8. The use according to claim 7, characterized in that The wheat germ agglutinin is used at a concentration of 0.36-0.45 g / L.

9. A method for plasma separation, characterized in that: The following steps are involved: Blood is mixed with wheat germ agglutinin and agglutinated; Filtering the blood after the coagulation treatment using the plasma filtration composite material according to claim 1 or 2, the plasma filter according to claim 3, or the plasma separation device according to claim 4 or 5; The filtered fractions were collected to obtain plasma.

10. The method according to claim 9, characterized in that The wheat germ agglutinin is used at a concentration of 0.36-0.45 g / L.

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