Antibody protective agent and blood collection tube

Abstract

The present invention provides an antibody protective agent, including a coagulant, a protective agent and an adsorbent material, wherein the protective agent includes thimerosal, urea, arginine, poloxamer, β-cyclodextrin, trehalose, copper sulfate, bovine serum albumin and a buffer. Compared with conventional coagulant blood collection tubes, which need to be left to stand for 30 minutes after blood collection to allow the blood to fully coagulate, and the centrifugal conditions are 3000 to 3500 rpm and centrifuged for 10 minutes to complete the separation of serum, the blood collection tubes in the present invention can not only effectively protect the antibodies in the serum and adsorb the interfering substances in the blood that affect the antibody detection, but also the serum separation operation is simple and fast, and high-purity serum can be obtained after centrifugation, which significantly shortens the time from blood sample collection to serum acquisition, and the entire serum separation process only takes 8 to 10 minutes. The present invention also provides a blood collection tube.

Description

Technical Field

[0001] The invention belongs to the technical field of blood testing, and relates to a storage container for serum antibodies in a whole blood sample, and in particular to an antibody protective agent and a vacuum blood collection tube composed of the antibody protective agent. Background Art

[0002] In order to meet the predetermined clinical test requirements, it is necessary to add reagents suitable for blood preservation or capable of reacting with blood samples into the vacuum blood collection tube. According to the type of additives, they are mainly divided into two categories: coagulants and anticoagulants. For some emergency biochemical tests or rapid serum immunological screening of infectious diseases, it is necessary to add coagulants into the blood collection tube to shorten the blood coagulation time. Coagulants can activate fibrinase, convert soluble fibrin into insoluble fibrin aggregates, and then form stable fibrin clots, accelerating blood coagulation and serum precipitation.

[0003] Some blood collection tubes containing coagulants will have additional separation gel added to form a barrier in the tube and completely separate the solid components (blood cells) and liquid components (serum). The specific gravity of the added separation gel is about 1.05, the specific gravity of serum is about 1.02, and the specific gravity of blood cells is about 1.08. When the separation gel and coagulated blood are centrifuged in the same blood collection tube, the blood clots heavier than the separation gel will move to the bottom of the tube, and the separation gel will reverse, forming a stacking order of blood cells / separation gel / serum from bottom to top. It is convenient for testers to extract the top layer of serum from the tube for downstream testing.

[0004] Currently, the antibody detection after conventional vaccine injection (such as hepatitis B, hepatitis C, etc.) and after infectious disease infection (such as new coronavirus, measles, AIDS, tuberculosis, etc.) uses the above-mentioned common coagulant / coagulant + separation gel blood collection tube. In the clinical testing process, due to incomplete blood coagulation and poor separation gel performance, fibrin will remain in the serum, resulting in errors in downstream test results. Moreover, the existing technologies are all applied to antibody protection in vitro, and the preservation and application of antibodies in blood samples such as serum are not involved. Summary of the invention

[0005] In view of this, the object of the present invention is to provide an antibody protective agent and a blood collection tube. The antibody protective agent provided by the present invention can promote rapid blood coagulation, protect antibody activity, and adsorb interfering substances in the blood that affect antibody detection, so as to obtain high-quality serum conveniently and quickly.

[0006] The present invention provides an antibody protective agent, comprising: a coagulant, a protective agent and an adsorption material;

[0007] The protective agent comprises: thimerosal, urea, arginine, poloxamer, beta-cyclodextrin, trehalose, copper sulfate, bovine serum albumin and buffer.

[0008] Preferably, the weight proportion of the coagulant is 5 to 50 parts;

[0009] The weight proportion of the thimerosal is 1 to 20 parts;

[0010] The weight proportion of the urea is 10 to 30 parts;

[0011] The weight proportion of the arginine is 10 to 40 parts;

[0012] The weight proportion of the poloxamer is 10 to 30 parts;

[0013] The weight proportion of the β-cyclodextrin is 1 to 30 parts;

[0014] The weight proportion of the trehalose is 1 to 20 parts;

[0015] The weight proportion of the copper sulfate is 1 to 10 parts;

[0016] The weight proportion of the bovine serum albumin is 1 to 20 parts;

[0017] The weight proportion of the buffer solution is 20 to 60 parts;

[0018] The weight proportion of the adsorbent material is 1 to 20 parts.

[0019] Preferably, the coagulant is silica powder.

[0020] Preferably, the particle size of the coagulant is 5 to 500 nm.

[0021] Preferably, the buffer is Tris-HCl buffer.

[0022] Preferably, the adsorption material is a resin or microspheres;

[0023] Preferably, the adsorption material is a specific adsorption material, and the material of the adsorption material is selected from one or more of polystyrene, polymethyl methacrylate, silica, a copolymer of styrene and carboxylic acid monomers, and agarose gel. Furthermore, the specific adsorption material can specifically adsorb colored groups such as bilirubin and heme in the blood. Furthermore, the specific adsorption material can specifically adsorb biotin in the blood. Furthermore, the specific adsorption material can specifically adsorb divalent magnesium ions in the blood.

[0024] Preferably, the mass percentage of the adsorption material in the antibody protective agent is 3% to 16.8%.

[0025] The present invention provides a blood collection tube, comprising:

[0026] The antibody protective agent and separation gel described in the above technical solution.

[0027] Preferably, the antibody protective agent and separation gel are arranged in the safety cap, rubber plug, tube body and / or tube interior of the blood collection tube.

[0028] After the blood sample is collected by a conventional blood collection tube, it needs to be left to stand for 30 minutes to allow the blood to fully coagulate and separate the serum. The blood collection tube of the present invention contains a nano-scale coagulant, and the specific surface area of ​​the coagulant is in the range of 250 to 350 m 2 / g, compared with conventional micron-level coagulants, the specific surface area has increased by 160 to 230 times, which can significantly increase the dispersion and the contact rate between the coagulant and fibrinogen in the blood, and can promote the rapid and efficient conversion of fibrinogen. The blood coagulation time is shortened to 1 / 10 of the original, and the coagulation effect is greatly improved. Conventional coagulant blood collection tubes need to be inverted and mixed 4 to 6 times after collecting venous blood. The blood collection tubes of the present invention only need to be inverted and mixed 2 to 3 times, and the blood can be completely coagulated within 2 to 3 minutes. There is no need to stand and wait, and it can be directly centrifuged. Compared with conventional coagulant blood collection tubes, the present invention significantly shortens the blood coagulation time, can improve the processing efficiency of clinical diagnostic samples, increase the sample processing volume, and increase the number of sample processing per unit time. The number of samples processed is increased by 5 to 10 times, and the sample processing efficiency is significantly improved.

[0029] Conventional blood collection tubes need to be left to stand for 30 minutes after blood collection before being placed in a centrifuge at 3000-3500 rpm for 8-10 minutes to separate the serum. Due to incomplete coagulation and poor separation gel performance, visible fibrin is often found in the serum, resulting in delays in downstream testing or errors in test results. The blood collection tubes of the present invention can separate the serum by centrifugation at 4000 rpm for 8 minutes.

[0030] The antibody protective agent is added to the blood collection tube of the present invention. During the inversion mixing and centrifugation of the blood collection tube, the antibody protective agent in the tube can effectively protect the concentration and activity of the antibody in the blood and increase the stability of the antibody. The surface of the specific adsorption material can specifically adsorb colored groups such as bilirubin and hemoglobin in the serum, and can also adsorb biotin and divalent magnesium ions, so that the interference substances in the blood can be effectively removed without affecting the composition of the serum and the subsequent antibody detection reaction. The adsorption material can be reversed with the separation gel during the centrifugation process and settled to the bottom layer of the blood collection tube, and will not remain in the serum, and will not affect the natural state of the serum. After the centrifugation, high-quality serum can be obtained. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The results are the test results of the antibody protection effect of the blood collection tubes prepared in the examples and comparative examples of the present invention. DETAILED DESCRIPTION

[0032] The technical solutions in the embodiments of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments in the present invention, all other examples improved or modified by ordinary technicians in this field belong to the scope of protection of the present invention. It should be understood that the embodiments of the present invention are only used to illustrate the technical effects of the present invention, rather than to limit the scope of protection of the present invention. In the embodiments, the methods used are conventional methods unless otherwise specified.

[0033] The present invention provides an antibody protective agent, comprising: a coagulant, a protective agent and an adsorption material;

[0034] The protective agent comprises: thimerosal, urea, arginine, poloxamer, beta-cyclodextrin, trehalose, copper sulfate, bovine serum albumin and buffer.

[0035] In the present invention, the antibody refers to a protein with protective effect produced by the body due to the stimulation of antigens. In the present invention, the antibody preferably includes: serum antibodies, hormones or polypeptides.

[0036] In the present invention, the weight proportion of the coagulant is preferably 5 to 50 parts, more preferably 7 to 43 parts, more preferably 10 to 40 parts, more preferably 15 to 35 parts, more preferably 20 to 30 parts, and most preferably 7 parts or 25 parts.

[0037] In the present invention, the coagulant is preferably silica powder; the particle size of the coagulant is preferably nanoscale, the particle size of the coagulant is preferably 5-500nm, more preferably 10-100nm, more preferably 15-50nm, more preferably 15-30nm, and most preferably 15nm.

[0038] In the present invention, the weight parts of the thimerosal are preferably 1 to 20 parts, more preferably 1 to 12 parts, more preferably 3 to 10 parts, more preferably 3 to 8 parts, and most preferably 3 parts.

[0039] In the present invention, the weight parts of the urea are preferably 10 to 30 parts, more preferably 16 to 27 parts, more preferably 18 to 22 parts, and most preferably 18 parts or 20 parts.

[0040] In the present invention, the weight proportion of the arginine is preferably 10 to 40 parts, more preferably 22 to 38 parts, more preferably 26 to 32 parts, and more preferably 26 or 30 parts.

[0041] In the present invention, the weight proportion of the poloxamer is preferably 10 to 30 parts, more preferably 16 to 26 parts, more preferably 18 to 22 parts, and most preferably 16 parts or 20 parts.

[0042] In the present invention, the weight proportion of the β-cyclodextrin is preferably 10 to 30 parts, more preferably 16 to 24 parts, more preferably 18 to 22 parts, and most preferably 16 parts or 20 parts.

[0043] In the present invention, the weight proportion of the trehalose is preferably 10 to 20 parts, more preferably 8 to 18 parts, more preferably 12 to 16 parts, and most preferably 12 parts or 16 parts.

[0044] In the present invention, the weight parts of the copper sulfate are preferably 1 to 10 parts, more preferably 1.5 to 7 parts, more preferably 2 to 5 parts, more preferably 3 to 4 parts, and most preferably 1.5 parts.

[0045] In the present invention, the weight portion of the bovine serum albumin is 1 to 20 parts, more preferably 1 to 12 parts, more preferably 3 to 10 parts, and most preferably 3 parts.

[0046] In the present invention, the weight parts of the buffer solution is preferably 20 to 60 parts, more preferably 25 to 55 parts, more preferably 30 to 50 parts, more preferably 35 to 45 parts, and most preferably 40 parts.

[0047] In the present invention, the buffer is preferably Tris-HCl buffer; the concentration of the buffer is preferably 0.08-0.12 mol / L, more preferably 0.1 mol / L. In the present invention, the buffer is preferably prepared with water, more preferably deionized water.

[0048] Different from the antibody protective agent in the prior art, which can only maintain protein and cell stability and inhibit proteolytic enzymes from hydrolyzing proteins, in the antibody protective agent of the present invention, arginine can inhibit the formation of multimeric antibodies and ensure the normal function of antibodies; urea can prevent the non-specific binding of rheumatoid factor (RF) and eliminate its interference with the immune detection reaction; copper sulfate can block lysozyme to prevent it from connecting to IgG antibody (immunoglobulin G, IgG) and eliminate its interference with the immune detection reaction. Nano-scale silicon powder greatly increases the dispersion, can quickly react with fibrinogen, promote blood to fully and quickly coagulate, and eliminate the influence of residual fibrinogen; while the adsorbent material adsorbs and eliminates the interference factors (magnesium ions\biotin\bilirubin) of the immune detection reaction, it can be precipitated at the bottom of the tube by centrifugation and separated from the serum, so as to ensure that the main composition and concentration of the serum remain unchanged and do not interfere with subsequent various detection and analysis.

[0049] In the present invention, the weight proportion of the adsorbent material is preferably 1 to 20 parts, more preferably 2 to 15 parts, more preferably 3 to 10 parts, and most preferably 3 parts.

[0050] In the present invention, the adsorption material is preferably a resin or a microsphere; the material of the adsorption material is preferably selected from one or more of polystyrene, polymethyl methacrylate, silica, a copolymer of styrene and carboxylic acid monomers, and agarose gel; the adsorption material is preferably a specific adsorption material; the specific adsorption material can specifically adsorb colored groups such as bilirubin and heme in the blood, and further, the specific adsorption material can specifically adsorb biotin in the blood, and further, the specific adsorption material can specifically adsorb divalent magnesium ions in the blood.

[0051] In the present invention, the mass percentage of the adsorbent material in the antibody protective agent is preferably 1% to 16.8%, more preferably 2 to 12%, more preferably 3 to 10%, more preferably 3 to 5%, and most preferably 3%.

[0052] In the present invention, the pH value of the antibody protective agent is preferably consistent with the pH value of the serum, which can ensure the stability of the entire serum and avoid the precipitation of antibodies caused by changes in pH value.

[0053] In the present invention, the method for preparing the antibody protective agent preferably comprises:

[0054] Dissolve urea in a buffer solution, then sequentially add poloxamer, β-cyclodextrin, trehalose, bovine serum albumin, thimerosal, arginine and copper sulfate, and finally add a coagulant, mix, and then use a buffer solution to make up the volume to obtain a mixed solution;

[0055] The mixed solution is mixed with an adsorption material to obtain an antibody protective agent.

[0056] In the present invention, it is preferred that the mixed solution and the adsorption material are mixed before using the antibody protective agent.

[0057] In the present invention, the mixing is preferably performed on a magnetic stirrer.

[0058] In the present invention, the method for preparing the mixed solution more preferably comprises:

[0059] Accurately weigh 18 parts by weight of urea, 16 parts by weight of poloxamer and 16 parts by weight of β-cyclodextrin, 12 parts by weight of trehalose, 3 parts by weight of bovine serum albumin and 3 parts by weight of thimerosal, 26 parts by weight of arginine, 1.5 parts by weight of copper sulfate and 7 parts by weight of silicon dioxide powder for later use; prepare 1 L of 0.1 mol / L Tris-HCl buffer with deionized water, and adjust the pH value to 7.2-7.6 for later use.

[0060] Take a clean beaker and first dissolve 18 parts by weight of urea in a small amount of 0.1 mol / L Tris-HCl buffer, then add 16 parts by weight of poloxamer and 16 parts by weight of β-cyclodextrin, 12 parts by weight of trehalose, 3 parts by weight of bovine serum albumin and 3 parts by weight of thimerosal, 26 parts by weight of arginine, and 1.5 parts by weight of copper sulfate in sequence. After each component is completely dissolved, the next component can be added. The entire dissolution process is carried out on a magnetic stirrer, stirring and dissolving at a uniform speed. Finally, 7 parts by weight of silicon powder are slowly added to the above mixed solution, and stirring is maintained at a uniform speed for 20 to 30 minutes. The volume is adjusted to a 100 mL volumetric flask with 0.1 mol / L Tris-HCl buffer, and the pH value is adjusted to 7.2 to 7.6 to obtain a mixed solution.

[0061] In the present invention, the antibody protective agent is added to the resin before use and mixed before use; the resin does not adsorb antibodies and does not affect the solubility and stability of proteins in the solution. The surface of the adsorbent material can specifically adsorb colored groups such as bilirubin and heme in the serum, and can also adsorb biotin and divalent magnesium ions, which can effectively remove interfering substances in the blood without affecting the composition of the serum or the subsequent antibody detection reaction. The adsorbent material can be reversed with the separation gel during the centrifugation process and settled to the bottom layer of the blood collection tube, and will not remain in the serum, and will not affect the natural state of the serum. High-quality serum can be obtained after the centrifugation.

[0062] The present invention provides a blood collection tube, comprising:

[0063] The antibody protective agent described in the above technical solution;

[0064] Separating gel.

[0065] In the present invention, the relative specific gravity of the separation gel is preferably 1.040 to 1.065 g / cm 3 , more preferably 1.04 g / cm 3 The viscosity is preferably 100,000 to 300,000 centipa, more preferably 150,000 to 250,000 centipa, and most preferably 200,000 centipa. In the present invention, the separation gel is preferably a colorless transparent colloid, physiologically inert, insoluble in water, and has excellent thixotropic and isolating properties.

[0066] The present invention has no particular limitation on the type and source of the separation gel, and serum separation gels well known to those skilled in the art may be used.

[0067] In the present invention, the mass ratio of the antibody protective agent to the separation gel is preferably (2-8): (5-30), more preferably (3-6): (10-25), and most preferably (4-5): (15-20).

[0068] In the present invention, the antibody protective agent and separation gel are preferably disposed in the safety cap, rubber plug, tube body and / or tube interior of the blood collection tube.

[0069] In the present invention, the blood collection tube is preferably a vacuum blood collection tube, and the interior of the blood collection tube is preferably a vacuum.

[0070] In the present invention, the method for preparing the blood collection tube preferably comprises:

[0071] The separation gel is added into the blood collection tube by a gel adding machine, and then the antibody protective agent is added into the blood collection tube after centrifugal sedimentation, and the tube is dried at 28-37° C. for 30-120 seconds, and the rubber stopper and safety cap are assembled, vacuumized, packaged, and sterilized to obtain the blood collection tube.

[0072] In the present invention, the glue applying machine is preferably a vacuum glue applying machine.

[0073] In the present invention, the amount of separation gel added is preferably 0.6 to 1.2 g, more preferably 0.8 to 1 g, and most preferably 0.9 g.

[0074] In the present invention, the centrifugation is carried out in a centrifuge; the centrifugation speed is preferably 2000-4000rpm / min, more preferably 2500-3500rpm / min, and most preferably 3000rpm / min; the centrifugation time is preferably 1-10min, more preferably 4-6min, and most preferably 5min.

[0075] In the present invention, the method for preparing the blood collection tube more preferably comprises:

[0076] Use a vacuum gelator to add 0.6-1.2 g of separation gel into the blood collection tube, centrifuge at 2000-4000 rpm / min for 1-10 min, add the antibody protective agent into the blood collection tube, dry at 28-37° C. for 30-120 s, assemble the rubber stopper and safety cap, evacuate, package, and sterilize to obtain the blood collection tube.

[0077] After the blood sample is collected by a conventional blood collection tube, it needs to be left to stand for 30 minutes to allow the blood to fully coagulate and separate the serum. The blood collection tube of the present invention contains a nano-scale coagulant, and the specific surface area of ​​the coagulant is in the range of 250 to 350 m 2 / g, compared with conventional micron-level coagulants, the specific surface area has increased by 160 to 230 times, which can significantly increase the dispersion and the contact rate between the coagulant and fibrinogen in the blood, and can promote the rapid and efficient conversion of fibrinogen. The blood coagulation time is shortened to 1 / 10 of the original, and the coagulation effect is greatly improved. Conventional coagulant blood collection tubes need to be inverted and mixed 4 to 6 times after collecting venous blood. The blood collection tubes of the present invention only need to be inverted and mixed 2 to 3 times, and the blood can be completely coagulated within 2 to 3 minutes. There is no need to stand and wait, and it can be directly centrifuged. Compared with conventional coagulant blood collection tubes, the present invention significantly shortens the blood coagulation time, can improve the processing efficiency of clinical diagnostic samples, increase the sample processing volume, and increase the number of sample processing per unit time. The number of samples processed is increased by 5 to 10 times, and the sample processing efficiency is significantly improved.

[0078] Conventional blood collection tubes need to be left to stand for 30 minutes after blood collection before being put into a centrifuge at 3000-3500 rpm for 10 minutes to complete the separation of serum. Due to incomplete coagulation and poor separation gel performance, visible fibrin is often found in the serum, resulting in delays in downstream testing or errors in test results. The blood collection tubes in the present invention can complete the separation of serum by centrifugation at 4000 rpm for 8 minutes.

[0079] The blood collection tube of the present invention is added with an antibody protective agent. During the inversion mixing and centrifugation of the blood collection tube, the antibody protective agent in the tube can effectively protect the concentration of antibodies in the blood and increase the stability of the antibodies. The surface of the specific adsorption material can specifically adsorb colored groups such as bilirubin and hemoglobin in the serum and can also adsorb biotin, which can effectively remove interfering substances in the blood without affecting the composition of the serum and the subsequent antibody detection reaction. The adsorption material can be reversed with the separation gel during the centrifugation process and settled to the bottom layer of the blood collection tube, and will not remain in the serum, and will not affect the natural state of the serum. High-quality serum can be obtained after the centrifugation.

[0080] The Tris-HCl buffer used in the following examples of the present invention is CAS No. 1185-53-1; thimerosal is CAS No. 54-64-8; arginine is CAS No. 74-79-3; urea is CAS No. 57-13-6; silicon dioxide powder is CAS No. 7631-86-9, poloxamer is CAS No. 9003-11-6; β-cyclodextrin is CAS No. 7585-39-9; trehalose is CAS No. 99-20-7; bovine serum albumin is CAS No. 9048-46-8; and copper sulfate is CAS No. 7758-98-7.

[0081] Example 1

[0082] Prepare the antibody protective agent as follows:

[0083] Accurately weigh 18 parts by weight of urea, 16 parts by weight of poloxamer and 16 parts by weight of β-cyclodextrin, 12 parts by weight of trehalose, 3 parts by weight of bovine serum albumin and 3 parts by weight of thimerosal, 26 parts by weight of arginine, 1.5 parts by weight of copper sulfate and 7 parts by weight of silicon dioxide powder for later use; prepare 1 L of 0.1 mol / L Tris-HCl buffer with deionized water, and adjust the pH value to 7.2-7.6 for later use.

[0084] Take a clean beaker and dissolve 18 parts by weight of urea in a small amount of 0.1mol / L Tris-HCl buffer, then add 16 parts by weight of poloxamer and β-cyclodextrin, 12 parts by weight of trehalose, 3 parts by weight of bovine serum albumin and thimerosal, 26 parts by weight of arginine, and 1.5 parts by weight of copper sulfate in sequence. After each component is completely dissolved, the next component can be added. The entire dissolution process is carried out on a magnetic stirrer, stirring and dissolving at a uniform speed. Finally, slowly add 7 parts by weight of silicon powder to the above mixture, keep stirring at a uniform speed for 20 to 30 minutes, dilute to a 100mL volumetric flask with 0.1mol / L Tris-HCl buffer, and adjust the pH value to 7.2 to 7.6 to obtain an antibody protective agent. (The amount of Tris-HCl buffer used is 56 parts by weight)

[0085] Before use, the above antibody protective agent is added with 3 parts by weight of resin (the resin component is polystyrene), mixed and then used to obtain the antibody protective agent.

[0086] Prepare blood collection tubes according to the following method:

[0087] Use a vacuum gelator to add 0.9 g of serum separation gel into the blood collection tube, centrifuge at 3000 rpm / min for 5 min, add 50 μL of the antibody protective agent prepared above into the blood collection tube, dry at 35°C for 60 s, assemble the rubber stopper and safety cap, evacuate, package, and sterilize to obtain the blood collection tube.

[0088] Embodiments 2 to 6

[0089] An antibody protective agent and a vacuum blood collection tube were prepared according to the method of Example 1, which differed from Example 1 in that the antibody protective agent formula in Table 1 was used.

[0090] Comparative Examples 1-2

[0091] An antibody protective agent and a vacuum blood collection tube were prepared according to the method of Example 1, which differed from Example 1 in that the antibody protective agent formula in Table 1 was used.

[0092] Table 1 Antibody protective agent formulations in Examples 1 to 6 and Comparative Examples 1 to 2

[0093]

[0094] Performance Testing

[0095] The serum separation method is:

[0096] Taking out the blood collection tubes prepared in the embodiment and the comparative example;

[0097] For venous blood collection, gently invert the blood collection tube 2 to 3 times after blood collection;

[0098] No need to wait, just place the blood collection tube in a centrifuge at 4000 rpm for 8 minutes to separate the serum.

[0099] Blood coagulation time detection method:

[0100] Take out the blood collection tubes prepared by the embodiment and the comparative example, and take 30 tubes from each group;

[0101] For venous blood collection, gently invert the blood collection tube 2 to 3 times to mix it evenly.

[0102] 1 mL of blood was quickly taken out from each embodiment and comparative example into a 1.5 ml centrifuge tube, and each embodiment and comparative example was taken 3 times. The blood coagulation time was measured using a blood coagulation method. The centrifuge tube was tilted at an angle of 15° every 15 seconds to observe whether the liquid surface was tilted until the blood in each tube stopped flowing. The coagulation time (CT, the time required for complete coagulation of blood after it was removed from the body) was observed and recorded. The test results are shown in Table 2, which shows the blood coagulation time of each tube.

[0103] Table 2 Blood coagulation time of blood collection tubes prepared in the embodiments of the present invention and the comparative examples

[0104] Group Test conditions Solidification time Experimental Results Comparative Example 1 Physiological Saline At room temperature, during the solidification process, the tube was tilted every 15 seconds. 660s Delayed solidification Comparative Example 2 Ordinary Silicon Powder At room temperature, during the solidification process, the tube was tilted every 15 seconds. 430s Normal coagulation Example 2 At room temperature, during the solidification process, the tube was tilted every 15 seconds. 90s Accelerated solidification Example 3 At room temperature, during the solidification process, the tube was tilted every 15 seconds. 90s Accelerated solidification Example 4 At room temperature, during the solidification process, the tube was tilted every 15 seconds. 105s Accelerated solidification Example 5 At room temperature, during the solidification process, the tube was tilted every 15 seconds. 105s Accelerated solidification Example 6 At room temperature, during the solidification process, the tube was tilted every 15 seconds. 90s Accelerated solidification Example 1 At room temperature, during the solidification process, the tube was tilted every 15 seconds. 75s Accelerated solidification

[0105] According to the blood coagulation time test results in Table 2, compared with the comparative example, the blood complete coagulation time after blood collection prepared by the blood collection tube of the embodiment of the present invention is significantly shortened.

[0106] The effect of serum separation was tested as follows:

[0107] According to the requirements of the 4.4 serum separation tube fibrin wall project of the health industry standard of the People's Republic of China "WST 224-2018 Performance Verification of Vacuum Blood Collection Tubes", the test was conducted. One group of blood collection tubes prepared in the embodiment and the comparative example, 40 tubes were taken from each group;

[0108] The second group is the blood collection tubes prepared in the embodiment and the comparative example. 40 tubes are taken from each group. The test conditions are changed. After blood collection, the blood collection tubes are directly placed in a centrifuge at 4000 rpm for 8 minutes to complete the separation of serum without waiting. The presence of fibrin wall in the blood collection tubes of the two experimental groups is observed and recorded; the test results are shown in Table 3.

[0109] Table 3 Serum separation effects of blood collection tubes prepared in the embodiments of the present invention and the comparative examples

[0110]

[0111]

[0112] The fibrin adhesion test results in Table 3 show that, compared with the comparative example, the serum prepared using the blood collection tubes of the embodiments of the present invention has no fibrin adhesion, the serum quality is good, and the downstream detection will not be inhibited and the downstream detection effect will not be affected.

[0113] Antibody protection effect detection:

[0114] An equal amount of hepatitis B surface antibody (HBsAb) serum (lyophilized) standard substance (No. GBW09163) was added to the blood collection tubes prepared in the embodiment and the comparative example, and 5 mL of goat serum incubated at 36°C was added to each tube, and the serum addition time was controlled at 10 to 40 seconds (simulating the venous blood collection time). The tubes were placed at room temperature for 96 hours, and then each blood collection tube was placed in a centrifuge at 4000 rpm for 8 minutes (simulating the separation of serum). The antibody titer in each group of blood collection tubes was directly determined by ELISA method. The operation steps are as follows:

[0115] Coating: Dilute the coating antigen (hepatitis B surface antigen (HBsAg) serum (lyophilized) standard material (No. GBW09164)) 500 times with ELISA coating solution (pH 9.6), add 100 μL to each well of the ELISA plate, and incubate at 37°C for 2 h;

[0116] Wash the plate: Pour off the liquid, add ELISA washing solution 5 times using a plate washer, and pat dry with absorbent paper;

[0117] Blocking: add 200 μL of 1% skim milk powder to each well and incubate at 37°C for 1.5 h;

[0118] Wash the plate: Pour off the liquid, add ELISA washing solution 5 times using a plate washer, and pat dry with absorbent paper;

[0119] Add primary antibody: serum containing hepatitis B surface antibody after treatment in different groups, dilute in multiples, add 100 μL to each well, and incubate at 37°C for 1 h;

[0120] Wash the plate: Pour off the liquid, add ELISA washing solution 5 times using a plate washer, and pat dry with absorbent paper;

[0121] Add enzyme-labeled antibody: add horseradish peroxidase-goat anti-human IgG diluted 10,000 times with PBST, add 100 μL to each well, and incubate at 37°C for 1 h;

[0122] Wash the plate: Pour off the liquid, add ELISA washing solution 5 times using a plate washer, and pat dry with absorbent paper;

[0123] Display: Add 100 μL of freshly prepared color developing solution to each well and incubate at 37°C in the dark for 10-15 min;

[0124] Stop: add 50 μL stop solution to each well;

[0125] Absorbance detection: The absorbance value of each sample well was measured at a wavelength of 450nm using an enzyme-labeled instrument.

[0126] Test results such as Figure 1 As shown, Figure 1 The horizontal axis is the antibody dilution, 1 to 8 represent 1:250, 1:500, 1:1000, 1:2000, 1:4000, 1:8000, 1:16000, 1:32000 respectively; the vertical axis is the absorbance. Figure 1 It can be seen that the antibody preservation effect of the embodiment of the present invention is significantly better than that of the comparative example, and the antibody titer is higher after the antibody is stored at room temperature for 96 hours.

[0127] Compared with conventional procoagulant blood collection tubes, which need to be left to stand for 30 minutes to allow the blood to fully coagulate, centrifugal conditions of 3000-3500 rpm, and centrifugation for 10 minutes to complete serum separation, the entire serum separation time takes 40-45 minutes. The blood collection tube of the present invention can not only effectively protect the antibodies in the serum, but also the serum separation operation is simple and fast, and high-purity serum can be obtained after centrifugation, which significantly shortens the time from blood sample collection to serum acquisition, and the entire serum separation process only takes 8-10 minutes.

[0128] After the blood sample is collected by a conventional blood collection tube, it needs to be left to stand for 30 minutes to allow the blood to fully coagulate and separate the serum. The blood collection tube of the present invention contains a nano-scale coagulant, and the specific surface area of ​​the coagulant is in the range of 250 to 350 m 2 / g, compared with conventional micron-level coagulants, the specific surface area has increased by 160 to 230 times, which can significantly increase the dispersion and the contact rate between the coagulant and fibrinogen in the blood, and can promote the rapid and efficient conversion of fibrinogen. The blood coagulation time is shortened to 1 / 10 of the original, and the coagulation effect is greatly improved. Conventional coagulant blood collection tubes need to be inverted and mixed 4 to 6 times after collecting venous blood. The blood collection tubes of the present invention only need to be inverted and mixed 2 to 3 times, and the blood can be completely coagulated within 2 to 3 minutes. There is no need to stand and wait, and it can be directly centrifuged. Compared with conventional coagulant blood collection tubes, the present invention significantly shortens the blood coagulation time, can improve the processing efficiency of clinical diagnostic samples, increase the sample processing volume, and increase the number of sample processing per unit time. The number of samples processed is increased by 5 to 10 times, and the sample processing efficiency is significantly improved.

[0129] Conventional blood collection tubes need to be left to stand for 30 minutes after blood collection before being placed in a centrifuge at 3000-3500 rpm for 8-10 minutes to separate the serum. Due to incomplete coagulation and poor separation gel performance, visible fibrin is often found in the serum, resulting in delays in downstream testing or errors in test results. The blood collection tubes of the present invention can separate the serum by centrifugation at 4000 rpm for 8 minutes.

[0130] The blood collection tube of the present invention is added with an antibody protective agent. During the inversion mixing and centrifugation of the blood collection tube, the antibody protective agent in the tube can effectively protect the concentration of antibodies in the blood and increase the stability of the antibodies. The surface of the specific adsorption material can specifically adsorb colored groups such as bilirubin and hemoglobin in the serum, and can also adsorb biotin and divalent magnesium ions, so that the interference substances in the blood can be effectively removed without affecting the composition of the serum and the subsequent antibody detection reaction. The adsorption material can be reversed with the separation gel during the centrifugation process and settled to the bottom layer of the blood collection tube, and will not remain in the serum, and will not affect the natural state of the serum. After the centrifugation, high-quality serum can be obtained.

[0131] The above-mentioned specific embodiments are only specific embodiments of the antibody storage tube of the present invention, and are not intended to limit the protection scope of the present invention. The storage of other types of proteins besides serum antibodies, such as hormones and polypeptides, are also within the application scope of the present invention. That is, any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

[0132] The above is only a preferred embodiment of the present invention. It should be pointed out that a person skilled in the art can make several improvements and modifications without departing from the principle of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. An antibody protective agent, include: Coagulants, protective agents and adsorbent materials; The protective agent includes: Thimerosal, urea, arginine, poloxamer, beta-cyclodextrin, trehalose, copper sulfate, bovine serum albumin, and buffer; The weight proportion of the coagulant is 5 to 50 parts; The weight proportion of the thimerosal is 1 to 20 parts; The weight proportion of the urea is 10 to 30 parts; The weight proportion of the arginine is 10 to 40 parts; The weight proportion of the poloxamer is 10 to 30 parts; The weight proportion of the β-cyclodextrin is 1 to 30 parts; The weight proportion of the trehalose is 1 to 20 parts; The weight proportion of the copper sulfate is 1 to 10 parts; The weight proportion of the bovine serum albumin is 1 to 20 parts; The weight proportion of the buffer solution is 20 to 60 parts; The weight proportion of the adsorbent material is 1 to 20 parts; The coagulant is silicon dioxide powder; the particle size of the coagulant is 5 to 500 nm; The buffer is Tris-HCl buffer; The adsorption material is resin or microspheres; the material of the adsorption material is selected from one or more of polystyrene, polymethyl methacrylate, silicon dioxide, copolymer of styrene and carboxylic acid monomers, and agarose gel; the mass percentage of the adsorption material in the antibody protective agent is 3% to 16.8%.

2. A blood collection tube, include: The antibody protective agent and separation gel according to claim 1.

3. The blood collection tube according to claim 2, It is characterized in that The antibody protective agent and separation gel are arranged in the safety cap, rubber plug, tube body and / or the inside of the blood collection tube.

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