CN215005422U - Blood collection tube for removing heterophilic antibody in serum or plasma - Google Patents
Blood collection tube for removing heterophilic antibody in serum or plasma Download PDFInfo
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- CN215005422U CN215005422U CN202023055639.XU CN202023055639U CN215005422U CN 215005422 U CN215005422 U CN 215005422U CN 202023055639 U CN202023055639 U CN 202023055639U CN 215005422 U CN215005422 U CN 215005422U
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Abstract
The utility model provides a blood collection tube for removing heterophilic antibodies in serum or plasma, which comprises a blood collection tube body; a connecting carrier layer is arranged on the inner wall of the blood collection tube body; the surface of the connecting carrier layer is crosslinked with a heterophilic antibody directional adsorption material layer; or the blood collection tube body is filled with connecting carrier particles; the surface of the connecting carrier particle is crosslinked with a heterophilic antibody directional adsorption material layer. Compared with the prior art, the utility model discloses be provided with the connection carrier layer at the body inner wall or fill in the body and connect the carrier particle, it has the directional adsorption material of heterophilic antibody to cross on the connection carrier, the blood sample after the blood sampling is accomplished is through reversing the mixing, makes heterophilic antibody in the sample fully contact with the connection carrier, and the carrier that is attached to heterophilic antibody after the centrifugation can be in blood clot layer or tube bottom, is about to separate heterophilic antibody from the serum sample, has guaranteed the accuracy of follow-up detection.
Description
Technical Field
The utility model belongs to the technical field of the heparin tube, especially, relate to a clear away heparin tube of heterophilic antibody in serum or plasma.
Background
Heterophilic Antibodies (HA) are a class of immunoglobulins with sufficient titers of multiple specificities to bind relatively weakly to immunoglobulins of multiple species stimulated by known or unknown antigenic substances in humans.
The heterophilic antibodies are produced in humans by exposure to contaminated food, exposure to fresh milk that has not been sterilized at high temperatures, feeding or exposure to animals and injection of vaccines derived from animal tissues or serum, animal-derived monoclonal antibodies used in immunotherapy, and the like, during human life. However, studies have now found that the production of xenotropic antibodies in humans is primarily due to direct contact with pets or laboratory mice.
Heterophilic antibodies are human endogenous antibodies and include both types of natural antibodies and autoantibodies, with natural antibodies being the predominant type of antibody that interferes. Heterophilic antibodies have a molecular weight of about 160kD and a relatively stable chemical structure. Since xenotropic antibodies have a large molecular weight and cannot pass through the glomerular filtration membrane of the kidney, they are generally present only in human blood and not in urine and other body fluids.
Due to the lack of knowledge of heterophilic antibodies, the examiner neglects the interference of antigen-antibody-based immunoassays, resulting in the inability to ensure the specificity and accuracy of the detection results. Referring to fig. 1 and 2, fig. 1 is a schematic diagram illustrating the reason why heterophilic antibodies interfere with the sandwich principle immunoassay, and heterophilic antibodies in blood bind to Fc or f (ab) of the coated antibody and the labeled antibody to form an empty sandwich, which results in a higher measurement signal value (pseudoincreased), or heterophilic antibodies in blood bind to the coated antibody and the labeled antibody, respectively, and cannot form a sandwich, which results in a lower measurement signal value (pseudodecreased); FIG. 2 is a schematic diagram showing the cause of immunological detection based on the principle of heterophilic antibody interference competition method, in which heterophilic antibody in blood binds to the antibody, blocking the binding of labeled antigen, resulting in a lower measured signal value (pseudoincreased).
At present, clinical diagnosis increasingly depends on test results to ensure that patients can obtain accurate diagnosis and treatment, so that immunoassay detection in the test process is not interfered, and the guarantee of the accuracy of the test results is very important. However, since heterophilic antibodies have no definite immunogen and interference thereof affects every reaction process for marker detection by using the immune principle, scientists and inspectors are prompted to intensively study interference mechanisms and search methods for eliminating heterophilic antibody interference.
SUMMERY OF THE UTILITY MODEL
In view of this, the to-be-solved technical problem of the present invention is to provide a blood collection tube for removing heterophilic antibodies in serum or plasma, which can perform directional adsorption on heterophilic antibodies in a blood sample, thereby ensuring the accuracy of subsequent detection results.
The utility model provides a blood collection tube for removing heterophilic antibodies in serum or plasma, which comprises a blood collection tube body; a connecting carrier layer is arranged on the inner wall of the blood collection tube body; the surface of the connecting carrier layer is crosslinked with a heterophilic antibody directional adsorption material layer.
Preferably, the heterophilic antibody directed adsorption material layer is an animal immunoglobulin layer or an animal immunoglobulin fragment layer.
Preferably, the connecting carrier layer is a resin layer.
Preferably, the thickness of the connecting carrier layer is 0.01-0.1 mm.
Preferably, the blood collection tube further comprises a tube cover for sealing the tube body of the blood collection tube; and a sealing soft plug is embedded in the middle of the pipe cover.
The utility model also provides a blood collection tube for removing heterophilic antibodies in serum or plasma, which comprises a blood collection tube body; the blood collection tube body is filled with connecting carrier particles; the surface of the connecting carrier particle is crosslinked with a heterophilic antibody directional adsorption material layer.
Preferably, the linking carrier particles are resin particles.
Preferably, the linking carrier particles are spherical particles; the diameter of the connecting carrier particles is 0.3-1 mm.
Preferably, the amount of the carrier particles filled in the blood collection tube is 1/10 to 1/5 of the volume of the blood collection tube.
Preferably, the heterophilic antibody directed adsorption material layer is an animal immunoglobulin layer or an animal immunoglobulin fragment layer.
The utility model provides a blood collection tube for removing heterophilic antibodies in serum or plasma, which comprises a blood collection tube body; a connecting carrier layer is arranged on the inner wall of the blood collection tube body; the surface of the connecting carrier layer is crosslinked with a heterophilic antibody directional adsorption material layer; or the blood collection tube body is filled with connecting carrier particles; the surface of the connecting carrier particle is crosslinked with a heterophilic antibody directional adsorption material layer. Compared with the prior art, the utility model discloses be provided with the connection carrier layer at the body inner wall or fill in the body and connect the carrier particle, it has the directional adsorption material of heterophilic antibody to cross on the connection carrier, the blood sample after the blood sampling is accomplished is through reversing the mixing, makes heterophilic antibody in the sample fully contact with the connection carrier, and the carrier that is attached to heterophilic antibody after the centrifugation can be in blood clot layer or tube bottom, is about to separate heterophilic antibody from the serum sample, has guaranteed the accuracy of follow-up detection.
Drawings
FIG. 1 is a schematic diagram showing the cause of heterophilic antibody interference in sandwich principle immunoassay;
FIG. 2 is a schematic diagram showing the cause of immunodetection based on the principle of heterophilic antibody interference competition;
fig. 3 is a schematic structural diagram of the blood collection tube for removing heterophilic antibodies in serum or plasma according to the present invention;
fig. 4 is a schematic structural diagram of the blood collection tube for removing heterophilic antibodies in serum or plasma provided by the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.
The utility model provides a blood collection tube for removing heterophilic antibodies in serum or plasma, which comprises a blood collection tube body; a connecting carrier layer is arranged on the inner wall of the blood collection tube body; the surface of the connecting carrier layer is crosslinked with a heterophilic antibody directional adsorption material layer.
Referring to fig. 3, fig. 3 is the utility model provides a clear away structure schematic diagram of heterophilic antibody blood sampling tube in serum or plasma, wherein 1 is the tube cap, 2 is sealed soft stopper, 3 is the blood sampling tube body, 4 are connecting the carrier layer.
The utility model provides a blood sampling tube which comprises a blood sampling tube body; the blood collection tube body can be a plastic tube body or a glass tube body, and is not particularly limited.
A connecting carrier layer is arranged on the inner wall of the blood collection tube body; the thickness of the connecting carrier layer is preferably 0.01-0.1 mm; the connecting carrier layer is preferably a resin layer, more preferably a thermoplastic resin layer; the resin layer is preferably provided with a group having a chemical function (such as-OH, -SH, -NH)2-COOH) polymer material; or the surface of the resin layer is preferably provided with a polymer layer with chemical functional groups; so as to be cross-linked with the heterophilic antibody directional adsorption material layer through a chemical functional group.
The surface of the connecting carrier is crosslinked with a heterophilic antibody directional adsorption material layer; the heterophilic antibody directed adsorption material layer is preferably an animal immunoglobulin layer, an animal immunoglobulin fragment layer, or a mixed layer of an animal immunoglobulin and a globulin fragment (comprising one or a combination of two or more of Fc, F (ab '), and F (ab')2 fragments).
According to the utility model, the blood sampling tube preferably further comprises a tube cover for sealing the tube body of the blood sampling tube; a sealing soft plug is embedded in the middle of the pipe cover; the tube cover is preferably a plastic tube cover.
In the present invention, the blood collection tube for removing heterophilic antibodies in serum or plasma can be prepared according to the following method: uniformly mixing the resin and the polymer solution at high temperature, and spraying the mixture on the inner wall of the tube body of the blood collection tube, wherein the thickness of the mixture is 0.01-0.1 mm; and cooling, soaking the blood sampling tube in an activating agent solution (such as 10-100 mmol/L carbodiimide) for 1h, washing with purified water twice, adding an heterophilic antibody directional adsorption material solution into the blood sampling tube, reacting at room temperature for 3h, and carrying out amidation reaction on carboxyl on the surface of the coating and amino of the heterophilic antibody directional adsorption material so as to introduce the heterophilic antibody directional adsorption material on the surface of the resin.
The utility model also provides a blood collection tube for removing heterophilic antibodies in serum or plasma, which comprises a blood collection tube body; the blood collection tube body is filled with connecting carrier particles; the surface of the connecting carrier particle is crosslinked with a heterophilic antibody directional adsorption material layer.
Referring to fig. 4, fig. 4 is the structure diagram of the blood collection tube for removing heterophilic antibodies in serum or plasma provided by the utility model, wherein 1 is a tube cap, 2 is a sealing soft plug, 3 is a blood collection tube body, and 5 is a connecting carrier particle.
The utility model provides a blood sampling tube which comprises a blood sampling tube body; the blood collection tube body can be a plastic tube body or a glass tube body, and is not particularly limited.
The blood collection tube body is filled with connecting carrier particles; the filling amount of the carrier particles connected in the blood collection tube body is preferably 1/10-1/5 of the volume of the blood collection tube body, and more preferably 1/10-1/8; the linking carrier particles are preferably spherical particles; the diameter of the connecting carrier particles is preferably 0.3-1 mm; the linking carrier particles are preferably resin particles, more preferably thermoplastic resin particles, still more preferably beaded thermoplastic resin spheres; the surface of the attached carrier particles is preferably coated with a coating with chemically functional groups (e.g., -OH, -SH, -NH)2-COOH) to cross-link the layer of heterophilic antibody directed adsorbent material via chemical functional groups.
The surface of the connecting carrier particle is crosslinked with a heterophilic antibody directional adsorption material layer; the heterophilic antibody directed adsorption material layer is preferably an animal immunoglobulin layer, an animal immunoglobulin fragment layer, or a mixed layer of an animal immunoglobulin and a globulin fragment (comprising one or a combination of two or more of Fc, F (ab '), and F (ab')2 fragments).
According to the utility model, the blood sampling tube preferably further comprises a tube cover for sealing the tube body of the blood sampling tube; a sealing soft plug is embedded in the middle of the pipe cover; the tube cover is preferably a plastic tube cover.
In the present invention, the blood collection tube for removing heterophilic antibodies in serum or plasma can be prepared according to the following method: firstly, connecting carrier particles such as bead-shaped thermoplastic resin spheres are soaked in an activator solution (such as 10-100 mmol/L carbodiimide) with the solid content of free carboxyl of the resin being 2-4 times of that of the free carboxyl of the resin for 1 hour, the connecting carrier particles are transferred into purified water to be soaked for 5 minutes, then the connecting carrier particles are soaked in an heterophilic antibody directional adsorption material solution to react for 3 hours at room temperature, and the carboxyl on the surfaces of the connecting carrier particles and the amino of the heterophilic antibody directional adsorption material have amidation reaction, so that the heterophilic antibody directional adsorption material is introduced to the surfaces of the connecting carrier particles.
The blood collection tube body, the tube cover and the sealing cork of the sealing blood collection tube body are manufactured according to a conventional process, vacuum is formed inside the sealing cork, when the sealing cork is used, a blood collection needle is pierced from the cork 3, negative pressure in the tube is utilized to collect blood, the blood in the blood collection tube is fully and uniformly mixed after blood collection is completed, heterophilic antibodies in a specimen are fully contacted with a connecting carrier layer or connecting carrier particles 4 which are crosslinked with a heterophilic antibody directional adsorption material layer, and the mixture is incubated at 37 ℃ for 30min or at room temperature for 60 min. After centrifugation of serum, the heterophilic antibody is attached to the wall of the blood collection tube to attach to a carrier layer 4 such as a thermoplastic resin or adsorbed onto the carrier particles 4 filled in the tube body of the blood collection tube.
In actual production, the linking carrier and the heterophilic antibody directed adsorbing material may be the same or similar materials as long as they have the same function.
The utility model discloses be provided with the connection carrier layer at the body inner wall or fill in the body and connect the carrier particle, it has the directional adsorption material of heterophilic antibody to cross on the connection carrier, the blood sample after the blood sampling is accomplished is through reversing the mixing, makes heterophilic antibody in the sample and connection carrier fully contact, and the carrier that has heterophilic antibody after the centrifugation can be in blood clot layer or tube bottom, is about to separate heterophilic antibody from the serum sample, has guaranteed the accuracy of follow-up detection.
In order to further explain the present invention, the following embodiments are combined to describe the blood collection tube for removing heterophilic antibodies in serum or plasma in detail.
The reagents used in the following examples are all commercially available.
EXAMPLE 1 preparation of heterophilic antibody directed adsorbent Material
The utility model relates to a heterophilic antibody directional adsorption material is a mixture containing animal immunoglobulin or globulin fragments such as bovine IgG, mouse IgG, rabbit IgG, sheep IgG, dog IgG and the like.
And (3) globulin purification process:
buffer preparation: 0.15M NaCl and 20mM Na at pH 8.0 were prepared2HPO4As binding/washing buffer; preparing 0.1M citric acid with the pH value of 2.5-3.0 as an elution buffer solution; after the preparation of the prepared reagent is finished, the prepared reagent is filtered by using a 0.22 mu m filter membrane for standby.
Sample preparation: mixing the animal serum to be purified with the binding/washing buffer solution at a ratio of 1:1, and storing at 2-8 ℃ for later use.
Column and resin preparation: binding/washing buffer was added to the column beforehand, Protein A/G purification resin was transferred to the column using a pipette, the resin was precipitated and the buffer was drained from the column, and then equilibrated with five column volumes of binding/washing buffer.
Sample purification: adding the sample to the resin at a slow flow rate; eluting the non-specifically adsorbed antibody by using binding/washing buffer solution with the volume of 10-15 times of the column volume; eluting with an elution buffer solution with 5-10 times of column volume; and collecting the effluent liquid, namely the globulin to be purified.
After elution of the globulin was complete, the resin was washed sequentially with 3 column volumes of binding buffer, 5 column volumes of deionized water, and 5 column volumes of 20% ethanol. The resin is added with 20 percent ethanol with the same volume and is stored at the temperature of 2-8 ℃ without being frozen.
Regeneration of the column: the column is washed with 10 column volumes of elution buffer and equilibrated with 5 column volumes of binding/washing buffer for reuse.
Mixing the purified animal globulins, or performing enzymolysis on the globulins to obtain fragments such as Fc, F (ab')2 and the like, mixing, wherein the concentration of each globulins is 0.5-10 mg/mL, and using the mixture as an allophilic antibody directional adsorption material.
EXAMPLE 2 preparation of serum-collecting blood vessel having thermoplastic resin coated on inner wall thereof
As shown in fig. 3, the utility model discloses the cross-linking has blood sampling tube of heterophilic antibody directional adsorption material, including the blood sampling tube body 3 that has sealed plastic tube lid 1, at plastic tube lid 1 middle part sealed the inlaying and be equipped with cork 2, the spraying has connection carrier layer 4 on the pipe wall in blood sampling tube body 3, uses the macromolecular layer thermoplastic resin that has chemical function group (carboxyl) in this embodiment as connecting carrier layer the thermoplastic resin surface cross-linking has heterophilic antibody directional adsorption material layer: firstly, uniformly mixing thermoplastic resin and high molecular solution at high temperature, uniformly spraying the mixture on the inner wall of a blood collection tube with the thickness of 0.01-0.1 mm, cooling the blood collection tube, soaking the blood collection tube in an activator solution (such as 10-100 mmol/L carbodiimide) for 1h, washing with purified water twice, adding an heterophilic antibody directional adsorption material solution into the blood collection tube, reacting for 3h at room temperature, and carrying out amidation reaction on carboxyl on the surface of a coating and amino of the heterophilic antibody directional adsorption material so as to introduce the heterophilic antibody directional adsorption material on the surface of the thermoplastic resin.
EXAMPLE 3 preparation of blood collecting tube containing beaded thermoplastic resin beads
As shown in fig. 4, the utility model discloses the cross-linking has blood sampling tube of heterophilic antibody directional adsorption material, including the blood sampling tube body 3 that has sealed plastic tube lid 1, it is equipped with soft stopper 2 to inlay in the middle part of plastic tube lid 1 is sealed, is filled with the connection carrier particle 5 that the diameter is 0.3 ~ 1.0mm in blood sampling tube body 3, this embodiment uses pearl-shaped thermoplastic resin ball as connecting carrier particle, and under normal condition, the filling amount of pearl-shaped thermoplastic resin ball is about 1/10 of body capacity, has heterophilic antibody directional adsorption material at pearl-shaped thermoplastic resin ball surface cross-linking: firstly, soaking a beaded thermoplastic resin ball in an activator solution (such as 10-100 mmol/L carbodiimide) with 2-4 times of resin free carboxyl solid content for 1h, transferring the resin ball into purified water to soak for 5min, then soaking the resin ball into an heterophilic antibody directional adsorption material solution, reacting for 3h at room temperature, and leading the heterophilic antibody directional adsorption material into the surface of the resin ball through amidation reaction of carboxyl on the surface of the resin ball and amino of the heterophilic antibody directional adsorption material; the reaction by-products with small particle size in the solution are screened out, and the solution can be placed into the blood collection tube body 3 for use after being washed by purified water.
The blood collection tube is manufactured according to a conventional process, vacuum is formed inside the blood collection tube, when the blood collection tube is used, the blood collection tube is penetrated through the soft plug 3, negative pressure in the tube is utilized to collect blood, the blood in the blood collection tube is fully and uniformly mixed after the blood collection is finished, the heterophilic antibodies in the sample are fully contacted with the beaded thermoplastic resin balls 4, and the beaded thermoplastic resin balls 4 attached with the heterophilic antibodies after centrifugation are positioned at a blood clot layer or the tube bottom.
Claims (8)
1. A blood collection tube for removing heterophilic antibodies in serum or plasma is characterized by comprising a blood collection tube body; a connecting carrier layer is arranged on the inner wall of the blood collection tube body; the surface of the connecting carrier layer is crosslinked with a heterophilic antibody directional adsorption material layer;
the connecting carrier layer is a resin layer; the resin layer is provided with a high polymer material with a chemical functional group or the surface of the resin layer is provided with a high polymer layer with a chemical functional group.
2. A blood collection tube according to claim 1, wherein the heterophilic antibody directed adsorption material layer is an animal immunoglobulin layer or an animal immunoglobulin fragment layer.
3. A blood collection tube according to claim 1, wherein the connecting carrier layer has a thickness of 0.01-0.1 mm.
4. A blood collection tube according to claim 1, further comprising a cap for sealing the tube body of the blood collection tube; and a sealing soft plug is embedded in the middle of the pipe cover.
5. A blood collection tube for removing heterophilic antibodies in serum or plasma is characterized by comprising a blood collection tube body; the blood collection tube body is filled with connecting carrier particles; the surface of the connecting carrier particle is crosslinked with a heterophilic antibody directional adsorption material layer;
the linking carrier particles are resin particles; the surface of the connecting carrier particle is coated with a macromolecule layer with chemical functional groups.
6. A blood collection tube according to claim 5, wherein the linking carrier particles are spherical particles; the diameter of the connecting carrier particles is 0.3-1 mm.
7. A blood collection tube according to claim 5, wherein the amount of the carrier particles attached to the tube body is 1/10 to 1/5 times the volume of the tube body.
8. A blood collection tube according to claim 5, wherein the heterophilic antibody directed adsorption material layer is an animal immunoglobulin layer or an animal immunoglobulin fragment layer.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202023055639.XU CN215005422U (en) | 2020-12-15 | 2020-12-15 | Blood collection tube for removing heterophilic antibody in serum or plasma |
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| CN202023055639.XU CN215005422U (en) | 2020-12-15 | 2020-12-15 | Blood collection tube for removing heterophilic antibody in serum or plasma |
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