Method for preparing special protein control products using blood product intermediate product components and special protein control products

Abstract

The application discloses a method for preparing special protein control products by using blood product intermediate product components and the special protein control products. The method provided in the application uses discarded intermediate product components in blood products as raw materials for preparing special proteins, which not only develops new raw materials with stable sources and controllable quality for preparing special proteins, but also opens up a new idea for recycling the intermediate products of blood products. The obtained products are guaranteed in biological safety, good in stability and high in cost performance.

Description

Technical Field

[0001] This invention relates to the field of biological waste recycling, and in particular to a method for preparing special protein control products using intermediate product components of blood products, and the special protein control products themselves. Background Technology

[0002] Blood products refer to therapeutic products derived from blood or plasma, such as red blood cells, platelets, plasma components, human serum albumin, human immunoglobulin products, and human coagulation factor products. The raw materials for these products are derived from normal human blood or plasma. Normal human blood or plasma is separated using a low-temperature ethanol process to obtain semi-finished or finished blood products. In the manufacturing process, intermediate products such as components I, II+III, or IV are produced in large quantities but have low recycling value and are often disposed of as waste. There is an urgent need in this field to explore the recycling value of these wastes.

[0003] Specialty proteins, also known as specific proteins, are many proteins found in human serum. These proteins originate from tissue cells and are functional proteins in serum, performing various functions. Many diseases can cause changes in serum protein levels. Proteins possess strong antigenicity, and antigen-antibody reactions can be used to detect proteins with specific antigenicity; hence, this is also called specific protein detection. Specialized protein tests include infectious marker proteins such as IgG, IgM, and IgA; rheumatic markers such as anti-streptolysin (ASO), rheumatoid factor (RF), and C-reactive protein (CRP); acute inflammatory markers such as CRP, α1-acid glycoprotein (AAG), haptoglobin (HPT), and α1-antitrypsin (AAT); nutritional status markers such as prealbumin (PA) and transferrin (TRF); and renal function markers such as α1-microglobulin (α1-MG) and β2-microglobulin (β2-MG). The detection of these specific proteins can assist doctors in diagnosis, for example, in the diagnosis of leukemia, multiple myeloma, autoimmune diseases, non-Hodgkin's lymphoma, rheumatic or rheumatoid diseases, infectious diseases, malnutrition, anemia, and glomerular and tubular dysfunction. To obtain stable and reliable diagnostic results, clinical laboratories need to establish a stable and reliable quality control system, and establishing a stable and reliable quality control system for specialty proteins depends on high-quality specialty protein control products.

[0004] However, currently, quality control materials for clinical specialty protein laboratory testing in China mainly come from imports or are non-standardized products temporarily prepared by laboratories using patient serum. Imported specialty protein controls are mostly single-item quality controls, with some being multi-item controls, but the number of test items is still limited. Quality control materials prepared using patient serum have low standardization and unstable quality; moreover, human serum and plasma are strictly controlled products, expensive, and limited in availability, making large-scale production impossible. Therefore, there is an urgent need in this field to develop a low-cost, commercially viable production process for multi-item composite specialty protein controls, providing technical and economic guarantees for quality control in specialty protein testing. Summary of the Invention

[0005] The purpose of this invention is to provide a method for preparing special protein control products using intermediate product components of blood products.

[0006] Another object of the present invention is to provide a special protein control product prepared by the above method.

[0007] To address the aforementioned technical problems, a first aspect of the present invention provides a method for preparing a special protein control product using intermediate components of blood products, the method comprising the steps of:

[0008] High-value specialty protein raw materials were extracted from components II and III and component IV, respectively; and

[0009] Special protein control products are formulated using the aforementioned high-value special protein raw materials;

[0010] The special proteins include IgG, IgM, IgA, complement C3, complement C4, anti-streptolysin (ASO), rheumatoid factor (RF), C-reactive protein (CRP), α1-microglobulin (α1-MG), β2-microglobulin (β2-MG), retinol-binding protein (RBP), transferrin (TRF), prealbumin (PA), α1-acid glycoprotein (AAG), and α1-antitrypsin (AAT).

[0011] In some preferred embodiments, the extraction of high-value specialty protein raw materials from components II+III and IV respectively includes the following steps:

[0012] High-value raw materials for extracting IgG, IgM, IgA, complement C3, complement C4, anti-streptolysin (ASO), rheumatoid factor (RF), C-reactive protein (CRP), α1-microglobulin (α1-MG), β2-microglobulin (β2-MG), and retinol-binding protein (RBP) from components II+III; and

[0013] High-value raw materials for extracting transferrin (TRF), prealbumin (PA), α1-acid glycoprotein (AAG), and α1-antitrypsin (AAT) from component IV.

[0014] In some preferred embodiments, the extraction of high-value specialty protein raw materials from components II+III and IV respectively includes the following steps:

[0015] Components II and III were pretreated, and then high-value raw materials containing IgG, IgM, IgA, complement C3, complement C4, anti-streptolysin (ASO), rheumatoid factor (RF), C-reactive protein (CRP), α1-microglobulin (α1-MG), β2-microglobulin (β2-MG), and retinol-binding protein (RBP) were extracted from the pretreated components II and III; and

[0016] Component IV was pretreated, and then high-value raw materials, including transferrin (TRF), prealbumin (PA), α1-acid glycoprotein (AAG), and α1-antitrypsin (AAT), were extracted from the pretreated component IV.

[0017] In some preferred embodiments, the pretreatment of components II+III includes the following steps:

[0018] After dissolving components II+III, they were subjected to degreasing and impurity removal treatments in sequence to obtain pretreated components II+III.

[0019] In some preferred embodiments, pretreatment of component IV includes the following steps:

[0020] After dissolving component IV, it was subjected to degreasing and impurity removal treatments in sequence to obtain pretreated component IV.

[0021] In some preferred embodiments, dissolving component II+III includes the following steps: dissolving component II+III in a Tris solution with a pH of 7-8, allowing it to stand to form a gel, breaking up the gel and centrifuging it, and taking the supernatant to obtain the dissolved component II+III.

[0022] In some preferred embodiments, dissolving component IV includes the following steps: dissolving component IV in a Tris solution with a pH of 7-8, allowing it to stand to form a gel, breaking up the gel and centrifuging it, and taking the supernatant to obtain the dissolved component IV.

[0023] In some preferred embodiments, the defatting process includes a sequential partial defatting process and a full defatting process; wherein the partial defatting process includes the step of removing low-density lipoprotein (LDL) from the dissolved component, and the full defatting process includes the step of removing high-density lipoprotein (HDL) from the dissolved component after the partial defatting process.

[0024] In some preferred embodiments, the semi-degreasing treatment includes the following steps: taking the dissolved component, adding dextran sulfate (DS) to a concentration of 0.1-1 g / L, then adding CaCl2 to a final concentration of 0.05-0.2 mol / L, allowing it to stand and centrifuging, separating the precipitate and taking the supernatant.

[0025] In some preferred embodiments, the full defatting process includes the following steps: adding dextran sulfate (DS) to the dissolved component after partial defatting treatment to a concentration of 3-7 g / L, then adding CaCl2 to a final concentration of 0.05-0.2 mol / L, allowing it to stand and centrifuging, separating the precipitate and taking the supernatant.

[0026] In some preferred embodiments, the impurity removal process includes the following steps: decalcifying the fully degreased component, then adding ammonium sulfate to a mass concentration of 20-25 wt%, allowing it to stand and centrifuging, separating the precipitate and taking the supernatant.

[0027] In some preferred embodiments, the calcium removal treatment includes the steps of: adding excess potassium oxalate to the fully defatted component, allowing it to stand and centrifuging it, separating the precipitate and taking the supernatant.

[0028] In some preferred embodiments, the extraction of high-value raw materials containing IgG, IgM, IgA, complement C3, complement C4, anti-streptolysin (ASO), rheumatoid factor (RF), C-reactive protein (CRP), α1-microglobulin (α1-MG), β2-microglobulin (β2-MG), and retinol-binding protein (RBP) from pretreated fractions II+III includes the following steps:

[0029] S1, take the pretreated components II+III, add ammonium sulfate to a concentration of 40-45 wt%, let stand and centrifuge, separate the precipitate and supernatant to obtain the first precipitate and the first supernatant. The first precipitate is a high-value raw material containing IgA, IgM, IgG, complement C3, complement C4, anti-streptolysin O (ASO), C-reactive protein (CRP), and rheumatoid factor (RF); and

[0030] S2, take the first supernatant and add solid ammonium sulfate to a final concentration of 78-85wt%, let stand and centrifuge, discard the supernatant to obtain the second precipitate, the second precipitate is a high-value raw material of α1-microglobulin (α1-MG), β2-microglobulin (β2-MG) and retinol-binding protein (RBP).

[0031] In some preferred embodiments, the extraction of high-value raw materials, including transferrin (TRF), prealbumin (PA), α1-acid glycoprotein (AAG), and α1-antitrypsin (AAT), from pretreated component IV includes the following steps:

[0032] S11, take the pretreated component IV and add solid ammonium sulfate to a concentration of 78-85 wt%, let stand and centrifuge, discard the supernatant to obtain the third precipitate, the third precipitate is a high-value raw material of transferrin (TRF), prealbumin (PA), α1-acid glycoprotein (AAG) and α1-antitrypsin (AAT).

[0033] In some preferred embodiments, the obtained high-value specialty protein raw material is subjected to dialysis. Preferably, the dialysate used in the dialysis process is bovine serum albumin solution.

[0034] In some preferred embodiments, the preparation of the special protein control product using the high-value special protein raw material includes the following steps:

[0035] The special high-value protein raw material is prepared to a preset concentration using a quality control diluent, and then freeze-dried.

[0036] In some preferred embodiments, the quality control diluent includes: a surfactant (preferably Tween-20), sodium chloride, phosphate buffer, a stabilizer (preferably a combination of maltose and trehalose), and a protectant (preferably bovine serum albumin).

[0037] In some preferred embodiments, the quality control diluent comprises the following components:

[0038] Protectant...1-5%;

[0039] Surfactant…0.1-1%;

[0040] Sodium chloride...0.5-3wt%;

[0041] Phosphate buffer…0.5-2M;

[0042] Stabilizer...1-5%.

[0043] In some preferred embodiments, the composition of the quality control diluent is as follows:

[0044] Bovine serum albumin...2%;

[0045] Surfactant…0.2%;

[0046] Sodium chloride…0.9 wt%;

[0047] Phosphate buffer…1M;

[0048] Maltose...2%; and

[0049] Trehalose...2%.

[0050] In a second aspect, the present invention provides a special protein control product prepared by the above method.

[0051] Compared with the prior art, the present invention has at least the following advantages:

[0052] (1) The method provided by the present invention uses waste intermediate product components from blood products as raw materials for the preparation of special proteins. This not only develops new raw materials with stable sources and controllable quality for the preparation of special proteins, but also opens up new ideas for the recycling of intermediate products from blood products. The resulting products have guaranteed biosafety, good stability, and high cost performance.

[0053] (2) In this invention, up to 15 special protein raw materials are extracted from waste intermediate blood product components, and after diluent optimization and concentration ratio exploration, they are prepared into stable composite special protein control products to meet the needs of patients for multiple tests in one visit.

[0054] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0055] Existing blood product manufacturing processes generate many intermediate components, which are often treated as biological waste with low recycling rates. Through extensive and in-depth research, the inventors have developed a method for preparing up to 15 kinds of specialty protein composite quality control products using intermediate components from blood products as raw materials. This not only provides a new way to recycle these intermediate components but also solves the problem of the scarcity of specialty protein control products. This invention is based on this.

[0056] Method for preparing characteristic protein control samples

[0057] This invention relates to a method for preparing a special protein control product using intermediate product components of blood products, comprising the steps of: extracting high-value special protein raw materials from components II+III and component IV respectively; and formulating the special protein control product using the obtained high-value special protein raw materials.

[0058] The special proteins involved in the high-value raw materials for special proteins prepared by the method of this invention include IgG, IgM, IgA, complement C3, complement C4, anti-streptolysin (ASO), rheumatoid factor (RF), C-reactive protein (CRP), α1-microglobulin (α1-MG), β2-microglobulin (β2-MG), retinol-binding protein (RBP), transferrin (TRF), prealbumin (PA), α1-acid glycoprotein (AAG), and α1-antitrypsin (AAT), covering almost all common clinical special protein test items.

[0059] In a preferred embodiment of the present invention, the extraction of high-value special protein raw materials from components II+III and IV respectively includes: high-value raw materials of IgG, IgM, IgA, complement C3, complement C4, anti-streptolysin (ASO), rheumatoid factor (RF), C-reactive protein (CRP), α1-microglobulin (α1-MG), β2-microglobulin (β2-MG) and retinol-binding protein (RBP) extracted from components II+III; and high-value raw materials of transferrin (TRF), prealbumin (PA), α1-acid glycoprotein (AAG) and α1-antitrypsin (AAT) extracted from component IV.

[0060] Before extracting high-value raw materials from components II+III or IV, pretreatment is preferably required to remove lipoproteins, other proteins, and other impurities to directly obtain relatively pure specialty proteins. In a preferred embodiment of the present invention, the pretreatment steps include: firstly, dissolving components II+III or IV, and then performing defatting and impurity removal treatments. Both components II+III and IV can be obtained from commercially available sources. Components II+III and IV are gel-like at room temperature. The step of dissolving components II+III or IV includes: dissolving the gel-like components II+III or IV in a buffer solution (usually Tris solution) with a pH of 7-8, allowing it to stand to form a gel, breaking up the gel and centrifuging it, and taking the supernatant to obtain the dissolved components II+III or IV.

[0061] The defatting process includes sequential partial and full defatting steps. Specifically, the dissolved components II+III or IV undergo partial defatting first, followed by full defatting. The main purpose of the partial defatting is to remove low-density lipoprotein (LDL). This partial defatting process includes the following steps: taking the dissolved components, adding dextran sulfate (DS) to a concentration of 0.1-1 g / L, then adding CaCl2 to a final concentration of 0.05-0.2 mol / L, allowing it to stand and centrifuging, separating the precipitate and collecting the supernatant. The precipitate is low-density lipoprotein (LDL), which can be recovered and used in desired applications. The main purpose of the full defatting process is to remove high-density lipoprotein (HDL). As a complete defatting process, it includes the following steps: adding dextran sulfate (DS) to the semi-defatted component to a concentration of 3-7 g / L, then adding CaCl2 to a final concentration of 0.05-0.2 mol / L, allowing it to stand and centrifuging, separating the precipitate and taking the supernatant. The precipitate contains more than 90% high-density lipoprotein (HDL), with the remainder being other proteins. The precipitate is recovered to remove other proteins, and the obtained high-density lipoprotein can be applied to the desired field.

[0062] The inventors discovered in their research that components II+III or IV, after defatting, still contain some impurities. The presence of these impurities increases the difficulty of subsequently extracting high-purity specialty proteins. Therefore, in a preferred embodiment of this invention, further impurity removal is required after defatting to remove impurities such as proteins. This includes the following steps: calcium removal treatment is performed on the defatted components, followed by the addition of ammonium sulfate to a concentration of 20-25 wt%, followed by standing and centrifugation. The precipitate is separated, and the supernatant is collected; this precipitate is the impurity protein. Excessive CaCl2 is added during defatting, resulting in a large amount of calcium ions within the components. Therefore, calcium removal treatment can be performed before protein removal to remove a large amount of calcium ions. The precipitate obtained in the calcium removal step is calcium oxalate; other precipitating agents can also be added to precipitate calcium ions.

[0063] The process involves taking pretreated fractions II+III or IV and extracting the target specific proteins. Specifically, the extraction of high-value raw materials from pretreated fractions II+III for IgG, IgM, IgA, complement C3, complement C4, anti-streptolysin (ASO), rheumatoid factor (RF), C-reactive protein (CRP), α1-microglobulin (α1-MG), β2-microglobulin (β2-MG), and retinol-binding protein (RBP) includes the following steps: S1, taking pretreated fractions II+III, adding ammonium sulfate to a concentration of 40-45 wt%, allowing to stand and centrifuging to separate the precipitate and... The supernatant yields a first precipitate and a first supernatant. The first precipitate contains high-value raw materials including IgA, IgM, IgG, complement C3, complement C4, anti-streptolysin O (ASO), C-reactive protein (CRP), and rheumatoid factor (RF); and S2. Solid ammonium sulfate is added to the first supernatant to a final concentration of 78-85 wt%, allowed to stand, and centrifuged. The supernatant is discarded to obtain a second precipitate, which contains high-value raw materials including α1-microglobulin (α1-MG), β2-microglobulin (β2-MG), and retinol-binding protein (RBP). The extraction of high-value raw materials containing transferrin (TRF), prealbumin (PA), α1-acid glycoprotein (AAG), and α1-antitrypsin (AAT) from pretreated component IV includes the following steps: S11, adding solid ammonium sulfate to pretreated component IV to a concentration of 78-85 wt%, allowing it to stand, centrifuging, and discarding the supernatant to obtain a third precipitate, which is the high-value raw material containing transferrin (TRF), prealbumin (PA), α1-acid glycoprotein (AAG), and α1-antitrypsin (AAT). In a preferred embodiment of the present invention, the obtained high-value raw materials of special proteins can be subjected to dialysis to remove ammonium sulfate. The dialysis method is a conventional technique in the art.

[0064] The high-value specialty protein raw materials extracted from components II+III and IV are compounded at a preset concentration. The preset concentration of the specialty protein compound quality control material can be set according to the needs of clinical testing. In a preferred embodiment of the present invention, the preset concentration is as follows:

[0065] Quality control level one

[0066] Components content IGG 9.97g / l IGA 1.88g / l IGM 0.95g / l C3 1.07g / l C4 0.225g ASO 117 IU / ml RF 28 IU / ml CRP 11mg / l α1-MG 19.33 mg / L β2-MG 2.1 mg / L RBP 22mg / l TRF 1.45g / l PA 171mg / l AAT 0.74g / l AAG 0.36g / l

[0067] Quality control level two

[0068]

[0069]

[0070] The formulation of the quality control diluent used in the compounding is as follows: protein protectant…1-5%; surfactant…0.1-1%; sodium chloride…0.5-3wt%; phosphate buffer…0.5-2M; and stabilizer…1-5%. The stabilizer is preferably a mixture of maltose and trehalose, the surfactant is preferably Tween-20, and the protectant is preferably bovine serum albumin. When the aforementioned quality control diluent is used with high-value specialty protein raw materials, the resulting product exhibits superior stability.

[0071] Furthermore, the obtained special protein complex quality control samples can be freeze-dried for easy storage. Freeze-drying can be performed using conventional techniques in this field.

[0072] This invention also relates to special protein control products prepared using the preparation method of this invention.

[0073] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the present invention is further described below in conjunction with specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight. Unless otherwise specified, the experimental materials and reagents used in the following embodiments are commercially available.

[0074] Unless otherwise specified, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. It should be noted that the terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the exemplary embodiments of this application.

[0075] Unless otherwise specified, the term “or” means the term “and / or” and is used interchangeably with the term “and / or”.

[0076] As used herein, including the appended claims, unless the context clearly indicates otherwise, the singular forms of words such as “an,” “a,” and “the” include their respective plural referents.

[0077] Example 1

[0078] In this embodiment, high-value raw materials including IGG, IGM, IGA, C3, C4, ASO, RF, CRP, α1-MG, β2-MG, and RBP were extracted from component II+III (purchased from Shanghai RAAS Blood Products Co., Ltd.). The specific steps are as follows:

[0079] (1) Component dissolution

[0080] Dissolve 1 kg of component II+III in 5 L of 0.02 M Tris pH 7.7 solution, stir and let stand overnight. The next day, break up the gel, centrifuge and filter, discard the precipitate and take the supernatant.

[0081] (2) Semi-degreasing treatment

[0082] Take 1L of the supernatant from step 1, place it on a stirrer and stir. Dissolve 0.5g of dextran sulfate (DS) in 5ml of purified water and slowly add it to the supernatant. After stirring for 5 minutes, add 2mol / L CaCl2 to a final concentration of 0.1mol / L. After stirring for 5 minutes, let it stand overnight and then centrifuge. Keep the precipitate as a high-value LDL raw material and take the supernatant.

[0083] (3) Complete degreasing treatment

[0084] Take 1L of the supernatant from step 2, place it on a stirrer and stir. Dissolve 5g of dextran sulfate (DS) in 50ml of purified water and slowly add it to the supernatant. After stirring for 5 minutes, add 2mol / L CaCl2 to a final concentration of 0.1mol / L. After stirring for 5 minutes, let it stand overnight and then centrifuge. Keep the precipitate as a high-value HDL raw material and take the supernatant.

[0085] (4) Impurity removal treatment

[0086] Take 1L of the supernatant from step 3, place it on a stirrer and stir, add 17g of K2C2O4, stir for 5 minutes and let stand overnight, centrifuge and discard the precipitate, take the supernatant, add solid ammonium sulfate to the supernatant to a final concentration of 22%, stir for 2 hours and let stand overnight, centrifuge and take the supernatant, the precipitate is impurities protein.

[0087] (5) Extract high values ​​of the five immune markers (IgG, IgM, IgA, C3, C4) and the three rheumatoid markers (ASO, RF, CRP).

[0088] Take the supernatant from step 4, add solid ammonium sulfate to a final concentration of 45%, stir for 2 hours and let stand overnight. After centrifugation, take the precipitate, which is the high value of the five immune items and three rheumatic items. Dialyze with PBS solution containing 0.2% bovine white blood cells until no ammonium sulfate is found, and keep the supernatant for later use.

[0089] (6) High values ​​of three parameters (α1-MG, β2-MG, RBP) in extracted urine

[0090] Take the supernatant from step 5, add solid ammonium sulfate to a final concentration of 80%, stir for 2 hours, let stand overnight, centrifuge, and take the precipitate, which is the high value of the three urine parameters. Dialyze with PBS solution containing 0.2% bovine leukopenia until no ammonium sulfate is found, and discard the supernatant.

[0091] (7) Measured values

[0092] Take the dialysis solutions from steps 5 and 6 and measure the concentration values ​​of each item using a biochemical analyzer.

[0093] Example 2

[0094] In this embodiment, high-value raw materials of TRF, PA, AAG, and AAT are obtained from the purification of component IV-4. The specific steps are as follows:

[0095] (1) Component dissolution

[0096] Dissolve 1 kg of component IV-4 in 5 L of 0.02 M Tris pH 7.7 solution, stir and let stand overnight. The next day, break up the gel, centrifuge and filter, discard the precipitate and take the supernatant.

[0097] (2) Semi-degreasing treatment

[0098] Take 1L of the supernatant from step 1, place it on a stirrer and stir. Dissolve 0.5g of dextran sulfate (DS) in 5ml of purified water and slowly add it to the supernatant. After stirring for 5 minutes, add 2mol / L CaCl2 to a final concentration of 0.1mol / L. After stirring for 5 minutes, let it stand overnight and then centrifuge. Keep the precipitate as a high-value LDL raw material and take the supernatant.

[0099] (3) Complete degreasing treatment

[0100] Take 1L of the supernatant from step 2, place it on a stirrer and stir. Dissolve 5g of dextran sulfate (DS) in 50ml of purified water and slowly add it to the supernatant. After stirring for 5 minutes, add 2mol / L CaCl2 to a final concentration of 0.1mol / L. After stirring for 5 minutes, let it stand overnight and then centrifuge. Keep the precipitate as a high-value HDL raw material and take the supernatant.

[0101] (4) Impurity removal treatment

[0102] Take 1L of the supernatant from step 3, add 17g of K2C2O4, stir for 5 minutes, let stand overnight, centrifuge to discard the precipitate, and take the supernatant.

[0103] (5) Extract high values ​​of TRF, PA, AAG and AAT.

[0104] Take the supernatant from step 4, add solid ammonium sulfate to a final concentration of 80%, stir for 2 hours, let stand overnight, centrifuge, and take the precipitate, which is the high value of TRF, PA, AAG, and AAT. Dialyze with PBS solution containing 0.2% bovine acetate until no ammonium sulfate is found, and discard the supernatant.

[0105] (6) Measured values

[0106] In step 5, the concentration values ​​of each item in the dialysis solution are measured using a biochemical analyzer.

[0107] Example 3

[0108] In this embodiment, the concentrations of 15 specific proteins obtained in Examples 1 and 2 were explored, and they were diluted to the target concentrations using a diluent with optimized formulation. The specific steps are as follows:

[0109] (1) Preparation of quality control diluent

[0110] Prepare special protein control diluents according to Table 1.

[0111] Table 1. Formulation of diluent for quality control products

[0112] Components content Bovine serum albumin 2％ Twain-20 0.2％ maltose 2％ Trehalose 2％ PBS 1M Sodium chloride 0.9％

[0113] (2) Use the quality control diluent to prepare the 15 special proteins to the concentrations shown in Table 2 below.

[0114] Table 2 Quality Control Level of Special Protein Control Products Formulation

[0115] Components content IGG 9.97g / l IGA 1.88g / l IGM 0.95g / l C3 1.07g / l C4 0.225g ASO 117 IU / ml RF 28 IU / ml CRP 11mg / l α1-MG 19.33 mg / L β2-MG 2.1 mg / L RBP 22mg / l TRF 1.45g / l PA 171mg / l AAT 0.74g / l AAG 0.36g / l

[0116] Quality control level two

[0117]

[0118]

[0119] Example 4

[0120] In this embodiment, the special protein control samples obtained in Example 3 were freeze-dried in a freeze dryer to obtain 15 special protein freeze-dried products. The specific freeze-drying process is shown in Table 3.

[0121] Table 3: Freeze-drying process

[0122] Temperature (°C) Duration (h) Temperature (°C) Duration (h) -30 12 0 2 -25 3 5 1 -20 2 10 1 -15 2 15 1 -10 2 20 1 -5 2 30 6

[0123] Those skilled in the art will understand that the above embodiments are specific examples of implementing the present invention, and in practical applications, various changes in form and detail may be made without departing from the spirit and scope of the present invention.

Claims

1. A method for preparing a special protein control product using blood product intermediate product components, characterized by, The method includes the following steps: High-value specialty protein raw materials were extracted from components II and III and component IV, respectively; and Special protein control products are formulated using the aforementioned high-value special protein raw materials; The special proteins include IgG, IgM, IgA, complement C3, complement C4, anti-streptolysin, rheumatoid factor, C-reactive protein, α1-microglobulin, β2-microglobulin, retinol-binding protein, transferrin, prealbumin, α1-acid glycoprotein, and α1-antitrypsin. The extraction of high-value specialty protein raw materials from components II+III and IV respectively includes the following steps: Components II and III were pretreated, and then high-value raw materials containing IgG, IgM, IgA, complement C3, complement C4, anti-streptolysin, rheumatoid factor, C-reactive protein, α1-microglobulin, β2-microglobulin, and retinol-binding protein were extracted from the pretreated components II and III; and Component IV was pretreated, and then high-value raw materials such as transferrin, prealbumin, α1-acid glycoprotein and α1-antitrypsin were extracted from the pretreated component IV. The extraction of high-value raw materials containing IgG, IgM, IgA, complement C3, complement C4, anti-streptolysin, rheumatoid factor, C-reactive protein, α1-microglobulin, β2-microglobulin, and retinol-binding protein from pretreated fractions II+III includes the following steps: S1, take the pretreated components II+III, add ammonium sulfate to a concentration of 40-45 wt%, let stand and centrifuge, separate the precipitate and supernatant to obtain the first precipitate and the first supernatant. The first precipitate contains IgA, IgM, IgG, complement C3, complement C4, anti-streptolysin, C-reactive protein and high-value raw materials of rheumatoid factor; and S2, take the first supernatant and add solid ammonium sulfate to a final concentration of 78-85 wt%, let stand and centrifuge, discard the supernatant to obtain the second precipitate, the second precipitate is a high-value raw material of α1-microglobulin, β2-microglobulin and retinol-binding protein; The extraction of high-value raw materials, including transferrin, prealbumin, α1-acid glycoprotein, and α1-antitrypsin, from pretreated component IV includes the following steps: S11, take the pretreated component IV, add solid ammonium sulfate to a concentration of 78-85 wt%, let stand and centrifuge, discard the supernatant to obtain the third precipitate, the third precipitate is a high-value raw material of transferrin, prealbumin, α1-acid glycoprotein and α1-antitrypsin.

2. The method of claim 1, wherein, The pretreatment of components II and III includes the following steps: After dissolving components II+III, they are sequentially subjected to degreasing and impurity removal treatments to obtain pretreated components II+III; and / or, Pretreatment of component IV includes the following steps: After dissolving component IV, it was subjected to degreasing and impurity removal treatments in sequence to obtain pretreated component IV.

3. The method of claim 2, wherein, Dissolving component II+III includes the following steps: dissolving component II+III in a Tris solution with a pH of 7-8, allowing it to stand to form a gel, breaking up the gel and centrifuging it, and collecting the supernatant to obtain the dissolved component II+III; and / or, Dissolving component IV involves the following steps: dissolving component IV in a Tris solution with a pH of 7-8, allowing it to stand to form a gel, breaking up the gel and centrifuging it, and taking the supernatant to obtain the dissolved component IV.

4. The method of claim 3, wherein, The defatting process includes a semi-defatting process and a full defatting process performed sequentially; wherein, the semi-defatting process includes the step of removing low-density lipoprotein from the dissolved component, and the full defatting process includes the step of removing high-density lipoprotein from the dissolved component after the semi-defatting process.

5. The method according to claim 4, characterized in that, The semi-degreasing treatment includes the following steps: take the dissolved component, add dextran sulfate to make its concentration 0.1-1 g / L, then add CaCl2 to the final concentration 0.05-0.2 mol / L, let it stand and centrifuge, separate the precipitate and take the supernatant. And / or, The complete defatting process includes the following steps: adding dextran sulfate to the dissolved component after partial defatting treatment to a concentration of 3-7 g / L, then adding CaCl2 to a final concentration of 0.05-0.2 mol / L, allowing it to stand and centrifuging, separating the precipitate and taking the supernatant.

6. The method according to claim 5, characterized in that, The impurity removal process includes the following steps: removing calcium from the fully degreased components, then adding ammonium sulfate to a mass concentration of 20-25 wt%, allowing it to stand and centrifuging, separating the precipitate and taking the supernatant.

7. Special protein control products prepared by the method according to any one of claims 1-6.

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