Gastric tumor composite quality control product and preparation method thereof

Abstract

This invention relates to the field of biotechnology, specifically to a composite quality control product for gastric tumor markers and its preparation method. The composite quality control product comprises gastric tumor marker antigens and a quality control buffer solution. The quality control buffer solution comprises 1-10% protein protectant, 0.01-3% surfactant, 2-20% thickening antioxidant, 0.5-500 mg / L protease inhibitor, 0.01-3% metal ion chelating agent, and 0.5-10% buffer matrix. The gastric tumor marker antigens include G17, PGII, and CA72-4. This invention, by addressing the interactions between G17, PGII, and CA72-4 antigens, achieves composite quality control for eight gastric tumor markers. This covers the quality control of four conventional gastric tumor detection reagents (PGI, PGII, G17, and CA72-4) and four broad-spectrum tumor detection reagents (AFP, CEA, CA19-9, and CA242), which is of significant importance for clinical gastric tumor screening.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a composite quality control product for gastric tumor markers and its preparation method. Background Technology

[0002] Quality control materials are used in the in vitro diagnostics (IVD) industry to control the quality of diagnostic reagents, playing a crucial role in the quality control of immunodiagnostics. Currently, the IVD industry primarily uses single-item quality control materials, typically bundled with a reagent kit for use as a single component. This means each control material can only be used for the quality control of one reagent. However, in clinical practice, multiple tests are usually conducted simultaneously. Single-item quality control materials require separate packaging, instrumentation, and selection for each test, which is cumbersome and time-consuming. Therefore, there is a need for multi-item quality control materials for combined testing, offering excellent quality control performance while facilitating convenient, rapid, time-saving, and labor-saving clinical applications. For these reasons, multi-item quality control materials have become the development trend, and both domestic and international IVD manufacturers highly value their development and application.

[0003] For clinical gastric tumor combined testing, the following indicators are generally tested together: alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), cancer antigen CA72-4 (CA72-4), pepsinogen I (PGI), pepsinogen II (PGII), carbohydrate antigen CA19-9 (CA19-9), carbohydrate antigen CA242 (CA242), and gastrin-17 (G17). However, current domestic and international IVD manufacturers' combined quality control products for gastric tumors cannot cover all of the above test items. This is mainly because adding too many antigens to a single buffer solution can lead to interactions between antigens, resulting in poor product stability. Therefore, it is currently impossible to achieve combined quality control of all eight gastric tumor markers. Summary of the Invention

[0004] This invention has found that in the quality control of gastric tumor markers, G17 antigen, PGII antigen and CA72-4 antigen are prone to mutual reaction, resulting in poor quality control stability and failure to achieve the purpose of quality control.

[0005] The purpose of this invention is to solve the problem of unstable composite quality control of gastric tumor markers.

[0006] Specifically as follows:

[0007] A gastric tumor marker composite quality control product includes a gastric tumor marker antigen and a quality control buffer. The quality control buffer comprises 1-10% protein protectant, 0.01-3% surfactant, 2-20% thickening antioxidant, 0.5-500 mg / L protease inhibitor, 0.01-3% metal ion chelating agent, and 0.5-10% buffer matrix. The gastric tumor marker antigen includes G17, PGII, and CA72-4.

[0008] The protein protectant described in this invention is mainly used to stabilize and protect the biological activity of antigens in solution and prevent non-specific binding;

[0009] The surfactant is used to increase the solubility of the protein;

[0010] The thickening antioxidant is used to reduce intermolecular motion, reduce oxidation of the buffer system, and increase the stability of the buffer system.

[0011] The protease inhibitor is used to inhibit proteases and protect proteins from degradation.

[0012] The metal ion chelating agent is used to chelate heavy metal ions to prevent them from damaging proteins.

[0013] The buffer matrix is ​​used to maintain the acid-base environment required for the reaction.

[0014] Preferably, the gastric tumor marker antigen also includes one or more of AFP, CEA, CA19-9, CA242, or PGI.

[0015] Preferably, the gastric tumor marker antigens include AFP, CEA, CA19-9, CA242, G17, PGI, PGII, and CA72-4;

[0016] Or the gastric tumor marker antigens may include G17, PGII, CA72-4 and PGI;

[0017] Or the gastric tumor marker antigens may include G17, PGII, CA72-4, AFP, CEA, CA19-9, and CA242;

[0018] Or the gastric tumor marker antigens mentioned may include G17, PGII, CA72-4, AFP, CEA and CA19-9;

[0019] Or the gastric tumor marker antigens may include G17, PGII, CA72-4, AFP, CEA and CA242.

[0020] The gastric tumor marker antigen of this invention is added for quality control purposes, so its concentration can be adjusted according to needs, and this invention does not limit it.

[0021] Preferably, the protein protectant is selected from casein, BSA, or HSA; the surfactant is selected from Tween 20, Tween 80, Brij 35, or Triton X-100; the thickening antioxidant is glycerol; the protease inhibitor is a pepsin inhibitor, aprotinin, or PMSF (serine protease); the metal ion chelating agent is selected from EDTA (ethylenediaminetetraacetic acid), NTA (aminotriacetic acid), or sodium gluconate; the buffer matrix is ​​selected from Tris, PBS, HEPES, or MES; more preferably, the EDTA is selected from EDTA·2Na or EDTA·2K.

[0022] Preferably, the concentration of the protein protectant is 1-3%, the concentration of the surfactant is 0.05-1%, the concentration of the glycerol is 10-15%, the concentration of the protease inhibitor is 0.5-500 mg / L, the concentration of the metal ion chelating agent is 0.01-1%, and the concentration of the buffer matrix is ​​0.5-1.5%.

[0023] 10. Preferably, the gastric tumor marker composite quality control product further includes a preservative, a protease inhibitor, a metal salt, or an antibacterial agent. Preferably, the preservative is selected from NaN3 or proclin300, the protease inhibitor is selected from pepsin inhibitor, aprotinin, or PMSF (serine protease), the metal salt is selected from sodium chloride or potassium chloride, and the antibacterial agent is selected from BND-10, PC950, or PC300.

[0024] More preferably, the concentration of the preservative is 0.01-3%, the concentration of the protease inhibitor is 0.5-500 mg / L, the concentration of the metal salt is 0.01-3%, and the concentration of the antibacterial agent is 0.01-3%.

[0025] More preferably, the pH of the gastric tumor marker composite quality control product is 6.8-7.2.

[0026] On the other hand, the present invention discloses a reagent or kit containing the aforementioned gastric tumor marker composite quality control product.

[0027] On the other hand, this invention discloses a method for preparing a composite quality control product for gastric tumor markers, comprising the following steps:

[0028] 1) Preparation of the quality control buffer solution;

[0029] 2) Preparation of the concentrated solution of the gastric tumor marker composite quality control product;

[0030] 3) Preparation of the gastric tumor marker composite quality control product.

[0031] On the other hand, this invention discloses the application of glycerol as a thickening and antioxidant in the aforementioned gastric tumor marker composite quality control product.

[0032] Beneficial effects

[0033] This invention discovers that the interaction between G17 antigen, PGII antigen, and CA72-4 antigen in the same buffer solution prevents the composite quality control of gastric tumor markers from achieving composite quality control of G17 antigen, PGII antigen, and CA72-4 antigen. This further limits the composite quality control of multiple gastric tumor markers. This invention, by resolving the interaction between G17 antigen, PGII antigen, and CA72-4 antigen, achieves composite quality control of eight gastric tumor markers. This covers the quality control of four conventional gastric tumor detection reagents (PGI, PGII, G17, and CA72-4) and four broad-spectrum tumor detection reagents (AFP, CEA, CA19-9, and CA242), which is of great significance for clinical gastric tumor screening. Detailed Implementation

[0034] To make the objectives, technical solutions, and advantages of the embodiments of this disclosure clearer, the technical solutions in the embodiments of this disclosure will be clearly and completely described below. Where specific conditions are not specified in the examples or embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0035] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. While any methods and materials similar to or equivalent to those described herein may be used in the practice or testing of formulations or unit doses herein, some methods and materials are described hereby. Unless otherwise stated, the techniques employed or considered herein are standard methods. Materials, methods, and examples are illustrative and not limiting in nature.

[0036] Preparation Example

[0037] ① Preparation of quality control buffer solution: Add each component of quality control buffer solution to pure water, mix thoroughly, and adjust the pH of the solution to 6.8-7.2 to obtain the buffer solution.

[0038] ② Preparation of concentrated gastric tumor marker composite quality control solution: The antigen was prepared into a high-concentration concentrated gastric tumor marker quality control solution using quality control buffer.

[0039] ③ Preparation of gastric tumor marker composite quality control: Low-value preparation: Calculate the corresponding amount of gastric tumor marker quality control concentrate based on the target concentration for the low value and add it to the quality control buffer. High-value preparation: Calculate the corresponding amount of gastric tumor marker quality control concentrate based on the target concentration for the high value and add it to the quality control buffer.

[0040] The final antigen quality control concentration is shown in the table below.

[0041] Table 1 Concentrations of various antigens in the gastric tumor marker composite quality control products

[0042] Project Name low value High value AFP 5ng / mL 250ng / mL CEA 3.5 ng / mL 50 ng / mL CA72-4 5U / mL 50U / mL PGI 15ng / mL 70ng / mL PGII 3ng / mL 30ng / mL CA19-9 20U / mL 75U / mL CA242 9U / mL 40U / mL G17 10 pmol / L 100 pmol / L

[0043] The quality control buffer solution includes a buffer matrix, protein protectant, surfactant, preservative, thickening antioxidant, protease inhibitor, metal salt, antibacterial agent, and metal ion chelating agent.

[0044] The buffer matrix is ​​selected from Tris, PBS, HEPES, or MES; the protein protectant is selected from casein, BSA, or HSA; the surfactant is selected from Tween 20, Tween 80, Brij 35, or Triton X-100; the thickening antioxidant is glycerol; the metal ion chelating agent is selected from EDTA (ethylenediaminetetraacetic acid), NTA (aminotriacetic acid), or sodium gluconate; the preservative is selected from NaN3 or proclin 300; the protease inhibitor is selected from pepsin inhibitor, aprotinin, or PMSF (serine protease); the metal salt is selected from sodium chloride or potassium chloride; and the antibacterial agent is selected from BND-10, PC950, or PC300.

[0045] In some embodiments, the concentration of the buffer matrix is ​​0.5%, 1%, 1.5%, 2%, 5%, or 10%.

[0046] In some embodiments, the concentration of the protein protectant is 1%, 3%, 5%, or 10%.

[0047] In some embodiments, the concentration of the surfactant is 0.01%, 0.05%, 0.1%, 1%, or 3%.

[0048] In some embodiments, the concentration of the thickening antioxidant is 2%, 5%, 10%, 15%, or 20%.

[0049] In some embodiments, the concentration of the metal ion chelating agent is 0.01%, 0.05%, 0.1%, 1%, or 3%.

[0050] In some embodiments, the concentration of the preservative is 0.01%, 0.05%, 0.1%, 1%, or 3%.

[0051] In some implementation examples, the concentrations of the protease inhibitors are 0.5 mg / L, 2 mg / L, 50 mg / L, 200 mg / L, and 500 mg / L;

[0052] In some embodiments, the concentration of the metal salt is 0.01%, 0.05%, 0.1%, 1%, or 3%.

[0053] In some embodiments, the concentration of the antibacterial agent is 0.01%, 0.05%, 0.1%, 1%, or 3%.

[0054] In some embodiments, the quality control buffer comprises 3% BSA, 1% Tween 20, 1% NaN3, 15% glycerol, 1% sodium chloride, 1% PC300, 0.5 mg / L pepsin inhibitor, 1% EDTA·2Na and 1.5% MES;

[0055] In some embodiments, the quality control buffer comprises 1% casein, 3% Triton X-100, 3% NaN3, 20% glycerol, 0.5 mg / L aprotinin, 3% potassium chloride, 3% BND-10, 3% EDTA·2K and 5% Tris;

[0056] In some embodiments, the quality control buffer comprises 1% casein, 0.1% Tween 80, 2 mg / L pepsin inhibitor, 10% glycerol, 0.1% NTA, and 2% PBS.

[0057] In some embodiments, the quality control buffer comprises 5% HSA, 3% Brij35, 50 mg / L PMSF, 20% glycerol, 0.1% proclin 300, 0.1% sodium chloride, 0.1% PC950, 0.05% EDTA·2Na, and 10% HEPES.

[0058] In some embodiments, the quality control buffer comprises 2% MES, 1% sodium chloride, 0.9% NaN3, 0.05% Tween 20, 1% BSA, 2% glycerol, 0.5 mg / L PMSF, 1% BND-10, and 1% EDTA·2Na;

[0059] In some embodiments, the quality control buffer comprises 3% HSA, 1% Tween 80, 2% PBS, 1% EDTA·2Na, 0.05% NaN3, 3% sodium chloride, 3% PC300, 50 mg / L aprotinin, and 2% glycerol.

[0060] In some embodiments, the quality control buffer comprises 2% Tris, 5% HSA, 0.05% Brij35, 200 mg / L PMSF, 5% glycerol, 0.05% EDTA·2K, 1% NaN3, 2% sodium chloride, and 1% PC950;

[0061] In some embodiments, the quality control buffer comprises 3% HEPES, 10% BSA, 3% Triton X-100, 20% glycerol, 200 mg / L pepsin inhibitor, 3% EDTA·2Na, 3% PC950, 1% sodium chloride, and 0.1% NaN3.

[0062] In some embodiments, the quality control buffer comprises 2% MES, 1% casein, 1% Tween 20, 0.01% NaN3, 0.05% EDTA·2K, 0.05% sodium chloride, 1% BND-10, 15% glycerol, and 500 mg / L aprotinin.

[0063] In some embodiments, the gastric tumor marker composite quality control product includes G17, PGII, CA72-4, 3% BSA, 1% Tween 20, 1% NaN3, 15% glycerol, 1% sodium chloride, 0.5 mg / L pepsin inhibitor, 1% PC300, 1% EDTA·2Na and 1.5% MES.

[0064] In some embodiments, the gastric tumor marker composite quality control product includes AFP, CEA, CA19-9, CA242, G17, PGI, PGII, CA72-4, 5% casein, 3% Triton X-100, 3% NaN3, 0.5 mg / L aprotinin, 20% glycerol, 3% potassium chloride, 3% BND-10, 3% EDTA·2K and 5% Tris.

[0065] In some embodiments, the gastric tumor marker composite quality control product includes G17, PGII, CA72-4, PGI, 2% MES, 1% sodium chloride, 0.9% NaN3, 0.05% Tween 20, 1% BSA, 2 mg / L pepsin inhibitor, 2% glycerol, 1% BND-10, and 1% EDTA·2Na;

[0066] In some embodiments, the gastric tumor marker composite quality control product includes G17, PGII, CA72-4, AFP, CEA, CA19-9, CA242, 3% HSA, 1% Tween 80, 2% PBS, 1% EDTA·2Na, 0.05% NaN3, 3% sodium chloride, 3% PC300, 50 mg / L PMSF, and 2% glycerol;

[0067] In some embodiments, the gastric tumor marker composite quality control product includes G17, PGII, CA72-4, AFP, CEA, CA19-9, 2% Tris, 5% HSA, 0.05% Brij35, 5% glycerol, 0.05% EDTA·2K, 1% NaN3, 2% sodium chloride, 50 mg / aprotinin, and 1% PC950;

[0068] In some embodiments, the gastric tumor marker composite quality control product includes G17, PGII, CA72-4, AFP, CA19-9, CEA, CA242, PGI, 3% HEPES, 10% BSA, 3% Triton X-100, 20% glycerol, 3% EDTA·2Na, 3% PC950, 1% sodium chloride, 200 mg / L PMSF, and 0.1% NaN3;

[0069] In some embodiments, the gastric tumor marker composite quality control product includes G17, PGII, CA72-4, AFP, CEA, CA242, 2% MES, 1% casein, 1% Tween 20, 0.01% NaN3, 0.05% EDTA·2K, 0.05% sodium chloride, 1% BND-10, 500 mg / L pepsin inhibitor, and 15% glycerol;

[0070] Example 1: Preparation and Validation of Gastric Tumor Marker Composite Quality Control Product I

[0071] The gastric tumor marker composite quality control product I is a composite quality control of G17 antigen, PGII antigen, and CA72-4 antigen.

[0072] The quality control buffer solution in this embodiment includes 3% BSA, 1% Tween 20, 1% NaN3, 15% glycerol, 1% sodium chloride, 1% PC300, 10 mg / L aprotinin, 1% EDTA·2Na, and 1.5% MES.

[0073] The specific preparation steps are as follows: ① Preparation of quality control buffer: Add each component of quality control buffer to pure water, mix thoroughly, and adjust the pH of the solution to 6.8-7.2 to obtain a quality control buffer composed of 3% BSA, 1% Tween 20, 1% NaN3, 15% glycerol, 1% sodium chloride, 1% PC300, 10 mg / L aprotinin, 1% EDTA·2Na and 1.5% MES;

[0074] ② Preparation of quality control concentrate: Prepare a CA72-4 quality control concentrate with a concentration of 2000 U / mL using the quality control buffer solution from step ①; prepare a PGII quality control concentrate with a concentration of 1000 ng / mL using the quality control buffer solution from step ①; prepare a G17 quality control concentrate with a concentration of 4000 pmol / L using the quality control buffer solution from step ①.

[0075] ③ Preparation of Gastric Tumor Marker Composite Quality Control Product I: Low-value preparation: Calculate the corresponding amount of CA72-4 quality control concentrate, PGII quality control concentrate, and G17 quality control concentrate according to the target concentration of the low value in Table 1, and add them to the quality control buffer in step ①; High-value preparation: Calculate the corresponding amount of CA72-4 quality control concentrate, PGII quality control concentrate, and G17 quality control concentrate according to the target concentration of the high value in Table 1, and add them to the quality control buffer in step ①.

[0076] The results are shown in Table 2. As can be seen from Table 2, when comparing the gastric tumor marker composite quality control product I placed at 37℃ for 10 days with that placed at 2-8℃ for 10 days (control group), the measured value deviation was within 10%, indicating that the quality control product has good stability.

[0077] Table 2. Data from accelerated stability and homogeneity experiments of the gastric tumor marker composite quality control material I.

[0078]

[0079]

[0080] Example 2: Preparation and Validation of Gastric Tumor Marker Composite Quality Control Product II

[0081] The gastric tumor marker composite quality control II is a composite quality control of AFP, CEA, CA19-9, CA242, G17, PGI, PGII and CA72-4.

[0082] The specific preparation steps in this embodiment are as follows: ① Preparation of quality control buffer: Add each component of quality control buffer to pure water, mix thoroughly, and adjust the pH of the solution to 6.8-7.2 to obtain a quality control buffer composed of 3% BSA, 1% Tween 20, 1% NaN3, 15% glycerol, 1% sodium chloride, 1% PC300, 10 mg / L aprotinin, 1% EDTA·2Na, and 1.5% MES;

[0083] ② Preparation of Quality Control Concentrates: Prepare a 10000 ng / mL AFP quality control concentrate using the quality control buffer from step ①; prepare a 5000 ng / mL CEA quality control concentrate using the quality control buffer from step ①; prepare a 2000 U / mL CA72-4 quality control concentrate using the quality control buffer from step ①; and prepare a 2000 ng / mL PGI quality control concentrate using the quality control buffer from step ①. Prepare a PGII quality control concentrate with a concentration of 1000 ng / mL using the quality control buffer from step ①. Prepare a CA19-9 quality control concentrate with a concentration of 5000 U / mL using the quality control buffer from step ①. Prepare a CA242 quality control concentrate with a concentration of 2000 U / mL using the quality control buffer from step ①. Prepare a G17 quality control concentrate with a concentration of 4000 pmol / L using the quality control buffer from step ①.

[0084] ③ Preparation of Gastric Tumor Marker Composite Quality Control II: Low Value Preparation: Add the concentrated AFP quality control solution, concentrated CEA quality control solution, concentrated CA72-4 quality control solution, concentrated PGI quality control solution, concentrated PGII quality control solution, concentrated CA19-9 quality control solution, concentrated CA242 quality control solution, and concentrated G17 quality control solution to the quality control buffer solution in step ①; High Value Preparation: Calculate the target concentration of the high value and add the corresponding amount of concentrated AFP quality control solution, concentrated CEA quality control solution, concentrated CA72-4 quality control solution, concentrated PGI quality control solution, concentrated PGII quality control solution, concentrated CA19-9 quality control solution, concentrated CA242 quality control solution, and concentrated G17 quality control solution to the quality control buffer solution in step ①.

[0085] ④ Evaluation method for accelerated stability and uniformity of quality control samples: The accelerated stability evaluation method is as follows: Quality control samples are divided into two groups and stored at two different temperatures; one group is stored at 37℃; the other group is stored at 2-8℃. Tests are performed after 3, 5, 7, and 10 days, with 2-3 replicates for each sample. The values ​​measured at 37℃ are compared with those measured at 2-8℃. The standard is: after 7 days at 37℃, the deviation between the measured values ​​and those at 2-8℃ should be ≤10%.

[0086] In this embodiment, the uniformity evaluation method is as follows: 10 or 15 quality control samples of the smallest packaging units in the same batch are randomly selected and randomly numbered 1 to 10 or 1 to 15. Each packaging unit is tested 3 times on the detection system. Considering the random variation of the measurement system due to factors such as time, the 3 measurements are performed in different orders, for example, 1, 3, 5, 7, 9, 2, 4, 6, 8, 10, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 2, 4, 6, 8, 10, 1, 3, 5, 7, 9. The test results are recorded, and F and S are calculated according to equations (1) to (11). bb S r and CV 瓶间 The results should meet the requirement that the inter-bottle CV should be ≤10%.

[0087]

[0088]

[0089] SS 瓶内 =SS 总和 -SS 瓶间 ……………………(3)

[0090]

[0091]

[0092]

[0093] υ1=α-1………………………………………… (7)

[0094] υ2=N-α………………………………………… (8)

[0095]

[0096]

[0097]

[0098] In the formula:

[0099] SS—Variance;

[0100] X i —Specify the i-th measurement value or calculation result of the parameter;

[0101] —Overall average;

[0102] n i —Number of repeated measurements for sample i;

[0103] X ij —The j-th result of sample i;

[0104] MS—mean square;

[0105] υ—degrees of freedom;

[0106] F-F test value;

[0107] n0—Number of valid measurements;

[0108] α—Number of samples drawn;

[0109] N—Total number of tests;

[0110] S bb —Standard deviation between bottles;

[0111] S r —Intra-bottle standard deviation (repeatability standard deviation).

[0112] When F≤1, with S r Replace S bb Calculate CV 瓶间 The results should meet the requirement that the inter-bottle CV should be ≤10%.

[0113] When F≤F 0.05 When (v1, v2), the test results show no significant difference in uniformity between bottles, and the calculation results are...

[0114] CV 瓶间The results should meet the requirement that the inter-bottle CV should be ≤10%.

[0115] When F > F 0.05 (v1,v2),S bb When the coefficient of variation is ≤0.3δ, the uniformity between bottles is considered to be good, and the calculated CV value is... 瓶间 The results should meet the requirement that the inter-bottle CV should be ≤10%.

[0116] When F > F 0.05 (v1,v2),S bb When the value is greater than 0.3δ, the uniformity between bottles is considered to be poor, which does not meet the requirement that the CV between bottles should be ≤10%.

[0117] Note 1: The sampling rule is that when the total number of samples is ≤500, the sampling size is 10, and when the total number of samples is >500, the sampling size is 15.

[0118] Note 2: δ is the target standard deviation.

[0119] In other embodiments, the uniformity evaluation method may be selected from the following methods:

[0120] Randomly select 10 or 15 bottles of quality control products from the same batch (the sampling rule is that when the total sample size is ≤500, the sampling size is 10; when the total sample size is >500, the sampling size is 15). Each bottle of quality control product is tested once using the matching detection system. Calculate the average value of the 10 or 15 test results according to formulas (1) to (2). Sum and standard deviation (S1); Additionally, randomly select one quality control sample from the above 10 or 15 samples and test it 10 or 15 times consecutively (if the quantity of one package is insufficient for 10 tests, only 5 consecutive tests are required), and calculate the average of the 10 or 15 test results. The standard deviation (S2) is calculated. The inter-bottle repeatability CV% is calculated according to formulas (3) to (4). The results should meet the requirement that the inter-bottle CV should be ≤10%.

[0121]

[0122]

[0123]

[0124]

[0125] When S1 < S2, let CV 瓶间 =0

[0126] In the formula:

[0127] -average value;

[0128] n - the number of measurements;

[0129] Xi - Specifies the i-th measurement value or calculation result of the parameter;

[0130] S - Standard deviation.

[0131] The results of accelerated stability and uniformity are shown in Table 3. It can be seen that when the gastric tumor marker composite quality control product II is placed at 37℃ for 10 days, compared with the control group placed at 2-8℃ for 10 days, the measured value deviation is within 10%, indicating that the quality control product has good stability.

[0132] It should be noted that the test values ​​may deviate from the theoretical concentrations in Table 1. This is because the actual test values ​​are within 30% of the theoretical concentrations, which is normal. Therefore, considering stability, the deviation of the measured values ​​only needs to be within 10%.

[0133] The homogeneity between and within bottles is shown in Table 4-5. The results show that the homogeneity between bottles of the eight items included in the gastric tumor marker composite quality control product II is within 4%, and the homogeneity within bottles of the eight items included in the gastric tumor marker composite quality control product II is within 3%, which meets the requirements of "YYT 1652-2019 General Technical Requirements for Quality Control Materials for In Vitro Diagnostic Reagents" (this industry standard requires that the homogeneity between quality control products should be ≤10%, and the homogeneity within bottles should be ≤8%).

[0134] Table 3. Data from accelerated stability and homogeneity tests of the gastric tumor marker composite quality control product II.

[0135]

[0136]

[0137] Table 4. Uniformity of Low Values ​​in Gastric Tumor Marker Composite Quality Control Material II

[0138]

[0139]

[0140]

[0141]

[0142] Table 5. Uniformity of High Values ​​in Gastric Tumor Marker Composite Quality Control Product II

[0143]

[0144]

[0145]

[0146]

[0147] Comparative Example 1: Modify the formulation of the quality control buffer.

[0148] This embodiment explores the impact of buffer formulation on quality control. The present invention creatively discovers that the interaction between G17 antigen, PGII antigen and CA72-4 antigen in the same buffer solution causes the gastric tumor marker composite quality control product to fail to achieve composite quality control of G17 antigen, PGII antigen and CA72-4 antigen, thereby further limiting the composite quality control of multiple gastric tumor markers (e.g., composite quality control of 8 gastric tumor markers).

[0149] Since the examples cannot be exhaustively listed, this example is prepared according to the method in Example 1. In this example, the stability of G17 antigen, PGII antigen and CA72-4 antigen in different buffer solutions is evaluated, as well as the stability of G17 antigen, PGII antigen and CA72-4 antigen plus the remaining AFP, CEA, CA19-9, CA242 and PGI in the same buffer solution is evaluated.

[0150] Table 6. Formulation of Quality Control Buffer

[0151]

[0152] Table 7. Stability of Formula 1 Gastric Tumor Marker Composite Quality Control Product I

[0153]

[0154]

[0155] Table 8. Stability of Formula 1 Gastric Tumor Marker Complex Quality Control Product II

[0156]

[0157]

[0158] Table 9. Stability of Formula 2 Gastric Tumor Marker Complex Quality Control Product I

[0159]

[0160]

[0161] Table 10 Stability of Formula 2 Gastric Tumor Marker Complex Quality Control Product II

[0162]

[0163]

[0164] Table 11. Stability of Formula 3 Gastric Tumor Marker Complex Quality Control Product I

[0165]

[0166]

[0167] Table 12 Stability of Formula 3 Gastric Tumor Marker Complex Quality Control Product II

[0168]

[0169]

[0170] Table 13 Stability of Formula 4 Gastric Tumor Marker Complex Quality Control Product I

[0171]

[0172]

[0173] Table 14 Stability of Formula 4 Gastric Tumor Marker Complex Quality Control Product II

[0174]

[0175]

[0176] Table 15 Stability of Formulation 5 Tumor Marker Composite Quality Control Product I

[0177]

[0178]

[0179] Table 16 Stability of Formula 5 Gastric Tumor Marker Complex Quality Control Product II

[0180]

[0181]

[0182] Table 17 Stability of Formulation 6 Tumor Marker Composite Quality Control Product I

[0183]

[0184]

[0185] Table 18 Stability of Formula 6 Gastric Tumor Marker Complex Quality Control Product II

[0186]

[0187]

[0188] Table 19 Stability of Formulation 7 Tumor Marker Composite Quality Control Product I

[0189]

[0190] Table 20 Stability of Formula 7 Gastric Tumor Marker Complex Quality Control Product II

[0191]

[0192]

[0193] The results are shown in Tables 7 to 20. As can be seen from Formulations 1-3, changing the concentration and type of components will not affect the stability and uniformity of the quality control products. Even removing preservatives, metal salts and antibacterial agents will not have a significant impact on stability. However, as can be seen from Formulations 4-6, the quality control products lacking protein protectants, surfactants, protease inhibitors, thickening antioxidants and metal ion chelating agents cannot maintain stable existence.

[0194] Meanwhile, the combined quality control of 3 antigens and 8 antigens showed the same trend, indicating that G17 antigen, PGII antigen and CA72-4 antigen have a decisive influence on the stability of the combined quality control. Solving the stability of the combined quality control of G17 antigen, PGII antigen and CA72-4 antigen can also solve the stability of more gastric tumor markers. Although this embodiment only lists the combined quality control of 8 antigens, it is not limited to 3 antigens and 8 antigens. Those skilled in the art can add one or more of AFP, CEA, CA19-9, CA242 or PGI to the G17 antigen, PGII antigen and CA72-4 antigen.

[0195] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A composite quality control product for gastric tumor markers, characterized in that, The gastric tumor marker composite quality control product includes gastric tumor marker antigen and quality control buffer. The quality control buffer includes 1-10% protein protectant, 0.01-3% surfactant, 2-20% thickening antioxidant, 0.01-3% metal ion chelating agent and 0.5-10% buffer matrix. The gastric tumor marker antigen includes G17, PGII and CA72-4.

2. The gastric tumor marker composite quality control product as described in claim 1, characterized in that, The gastric tumor marker antigens also include one or more of AFP, CEA, CA19-9, CA242, or PGI.

3. The gastric tumor marker composite quality control product as described in claim 2, characterized in that, The gastric tumor marker antigens include AFP, CEA, CA19-9, CA242, G17, PGI, PGII, and CA72-4.

4. The gastric tumor marker composite quality control product as described in claim 2, characterized in that, The gastric tumor marker antigens include G17, PGII, CA72-4, and PGI.

5. The gastric tumor marker composite quality control product as described in claim 2, characterized in that, The gastric tumor marker antigens include G17, PGII, CA72-4, AFP, CEA, CA19-9, and CA242.

6. The gastric tumor marker composite quality control product as described in claim 2, characterized in that, The gastric tumor marker antigens include G17, PGII, CA72-4, AFP, CEA, and CA19-9.

7. The gastric tumor marker composite quality control product as described in claim 2, characterized in that, The gastric tumor marker antigens include G17, PGII, CA72-4, AFP, CEA, and CA242.

8. The gastric tumor marker composite quality control product as described in any one of claims 1-7, characterized in that, The protein protectant is selected from casein, BSA, or HSA; the surfactant is selected from Tween 20, Tween 80, Brij 35, or Triton X-100; the thickening antioxidant is glycerol; the metal ion chelating agent is selected from EDTA (ethylenediaminetetraacetic acid), NTA (aminotriacetic acid), or sodium gluconate; and the buffer matrix is ​​selected from Tris, PBS, HEPES, or MES.

9. The gastric tumor marker composite quality control product as described in claim 8, characterized in that, The EDTA is selected from EDTA·2Na or EDTA·2K.

10. The gastric tumor marker composite quality control product as described in claim 8, characterized in that, The concentration of the protein protectant is 1-3%, the concentration of the surfactant is 0.05-1%, the concentration of the thickening antioxidant is 10-15%, the concentration of the metal ion chelating agent is 0.01-1%, and the concentration of the buffer matrix is ​​0.5-1.5%.

11. The gastric tumor marker composite quality control product as described in claim 8, characterized in that, The gastric tumor marker composite quality control product also includes preservatives, metal salts, or antibacterial agents.

12. The gastric tumor marker composite quality control product as described in claim 11, characterized in that, The preservative is selected from NaN3 or proclin300, the metal salt is selected from sodium chloride or potassium chloride, and the antibacterial agent is selected from BND-10, PC950 or PC300.

13. The gastric tumor marker composite quality control product as described in claim 12, characterized in that, The concentration of the preservative is 0.01-3%, the concentration of the metal salt is 0.01-3%, and the concentration of the antibacterial agent is 0.01-3%.

14. The gastric tumor marker composite quality control product as described in claim 8, characterized in that, The pH of the gastric tumor marker composite quality control product is 6.8-7.

2.

15. A reagent or kit, characterized in that, The reagent or kit contains the gastric tumor marker composite quality control product as described in any one of claims 1-14.

16. A method for preparing a gastric tumor marker composite quality control product as described in any one of claims 1-14, characterized in that, Includes the following steps: 1) Preparation of the quality control buffer solution as described in any one of claims 1-14; 2) Preparation of a concentrated solution of the gastric tumor marker composite quality control product as described in any one of claims 1-14; 3) Preparation of the gastric tumor marker composite quality control product as described in any one of claims 1-14.

17. The use of glycerol as a thickening and antioxidant in the gastric tumor marker complex quality control product as described in any one of claims 1-14.

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