Stable creatine kinase isoenzyme freeze-dried product and preparation method thereof

Abstract

The invention relates to the technical field of chemiluminescent in-vitro diagnostic reagents, in particular to a stable creatine kinase isoenzyme freeze-dried product and a preparation method thereof, and the stable creatine kinase isoenzyme freeze-dried product comprises the following components: 20-100mmol / L of a buffer solution with the pH value of 7.4 + / -0.2, 0.05-0.2% of a preservative, 0.05-0.2% of a preservative, 0.05-0.2% of The creatine kinase isoenzyme freeze-dried product is characterized in that the creatine kinase isoenzyme freeze-dried product comprises 2-20% of a protein stabilizer, 0.2-1% of a metal ion stabilizer, 0.01-0.03% of an inhibitor, 0.1-1% of amino acid, 5-15% of a sugar alcohol stabilizer, 10 mL / L of glycerol, 0.05-0.2% of a surfactant and 0-300 ng / mL of a creatine kinase isoenzyme antigen, and the creatine kinase isoenzyme freeze-dried product is stored under the sealing condition of 2-8 DEG C and has the validity period of 15 months; the creatine kinase isoenzyme freeze-dried calibrator prepared by the method has the advantages that the use period of the calibrator in an uncovered state in a matched instrument is 8 hours, the creatine kinase isoenzyme freeze-dried calibrator has good detection performance on the determination of creatine kinase isoenzyme (CK-MB), can be stored for 1 month at-20 DEG C after being redissolved and can be stored for 2 months at-20 DEG C, the performance is stable, the cost is low, the test result is accurate, the repeatability is good, and the problem that the existing creatine kinase isoenzyme freeze-dried calibrator is unstable in storage is solved.

Description

Technical Field

[0001] The invention relates to the technical field of chemiluminescent in vitro diagnostic reagents, and in particular to a stable creatine kinase isoenzyme freeze-dried product and a preparation method thereof. Background Art

[0002] At present, the determination of creatine kinase isoenzymes has a variety of auxiliary functions for clinical judgment: 1. Serum CK-MB and troponin levels may help to distinguish aortic dissection from myocardial infarction, unless the myocardial infarction is caused by dissection; 2. Acute myocardial infarction, myocarditis: creatine kinase MB isoenzyme [CK-MB]) is released into the bloodstream after myocardial cell necrosis, reaches a peak value in a short time, and then rapidly decays; 3. Acute coronary syndrome (ACS): Non-ST segment elevation MI (NSTEMI, subendocardial myocardial infarction): refers to myocardial necrosis without acute ST segment elevation (elevated myocardial markers troponin I / T and creatine kinase). The electrocardiogram may show ST segment depression, T wave inversion, or both. ST segment elevation MI (STEMI, transmural myocardial infarction): refers to myocardial necrosis accompanied by ST segment elevation (cannot be quickly reversed by nitroglycerin) or new left bundle branch block. Troponin I or troponin T and creatine kinase (CK) are elevated.

[0003] Common CK-MB on the market are all freeze-dried products, which need to be manually added with liquid for reconstitution when used. However, most freeze-dried products have poor effects, resulting in large differences between bottles, and poor stability after reconstitution, that is, the freeze-dried products of creatine kinase isoenzymes damage the antigen during freeze-drying and cannot be stored for a long time after reconstitution. Because creatine kinase isoenzymes are easily oxidized, some manufacturers add antioxidants to their freeze-dried products, resulting in a strong taste of the reconstituted product, which is not friendly to production configuration and clinical use.

[0004] More than 95% of the manufacturers of creatine kinase isoenzyme (CK-MB) kits currently on the market use freeze-dried calibrators. Even for freeze-dried calibrators, most have a shelf life of less than 12 months and are unstable in storage. Summary of the invention

[0005] The object of the present invention is to provide a stable creatine kinase isoenzyme freeze-dried product and a preparation method thereof, aiming to solve the problem of unstable storage of existing creatine kinase isoenzyme freeze-dried calibrators.

[0006] To achieve the above objectives, in a first aspect, the present invention provides a stable lyophilized product of creatine kinase isoenzyme, comprising the following components: 20-100 mmol / L of buffer solution with a pH of 7.4±0.2, 0.05-0.2% of preservative, 2-20% of protein stabilizer, 0.2%-1% of metal ion stabilizer, 0.01%-0.03% of inhibitor, 0.1%-1% of amino acid, 5%-15% of sugar alcohol stabilizer, 10 mL / L of glycerol, 0.05%-0.2% of surfactant and 0-300 ng / mL of creatine kinase isoenzyme antigen.

[0007] Wherein, the buffer comprises any one of MES buffer and a mixed buffer of MES and Tris, and the preservative comprises one or more of Proclin 300, NaN3 and gentamicin.

[0008] Among them, the metal ion stabilizer is NaCl, the protein stabilizer includes one or more of human serum albumin, bovine serum albumin, casein and animal serum, the amino acid includes any one of glycine and L-arginine, the inhibitor is phenylmethylsulfonyl fluoride, and the sugar alcohol stabilizer includes any two or more of trehalose, sucrose, sorbitol and dextran 500.

[0009] Wherein, the surfactant includes one or both of Triton 100 and Tween 20, and the enzyme isozyme antigen concentration is 0 ng / mL, 2 ng / mL, 10 ng / mL, 30 ng / mL, 60 ng / mL and 300 ng / mL.

[0010] In a second aspect, the present invention further provides a method for preparing a stable lyophilized product of creatine kinase isoenzyme, which is used to prepare the stable lyophilized product of creatine kinase isoenzyme as described in the first aspect, characterized in that it comprises the following steps:

[0011] Take a beaker and fill it with a pre-prepared volume of purified water, and add creatine kinase isoenzyme antigen and metal ion stabilizer to the beaker in sequence, stir and mix, adjust the pH, and obtain a primary mixed solution;

[0012] Adding a sugar alcohol stabilizer, an amino acid and a protein stabilizer to the primary mixed solution in sequence, stirring and dissolving, to obtain a secondary mixed solution;

[0013] Add sugar alcohol stabilizer, glycerol, inhibitor, surfactant and preservative to the secondary mixed solution in sequence, stir and dissolve, adjust the volume, filter with a filter membrane, and obtain creatine kinase isoenzyme freeze-dried product.

[0014] The invention discloses a stable creatine kinase isozyme freeze-dried product, comprising the following components: 20-100 mmol / L of buffer solution with a pH value of 7.4±0.2, 0.05-0.2% of preservative, 2-20% of protein stabilizer, 0.2%-1% of metal ion stabilizer, 0.01%-0.03% of inhibitor, 0.1%-1% of amino acid, 5%-15% of sugar alcohol stabilizer, 10 mL / L of glycerol, 0.05%-0.2% of surfactant and 0-300 ng / mL of creatine kinase isozyme antigen. The creatine kinase isozyme freeze-dried product is stored under a sealed condition of 2°C-8°C and has a validity period of 15 months. The product is placed in a matching container and placed in a container. The instrument has a shelf life of 8 hours when the lid is opened. It has good detection performance for the determination of creatine kinase isoenzymes (CK-MB) and can be stored at -20°C for 1 month and at -20°C for 2 months after reconstitution. It has stable performance, low cost, accurate test results, and good repeatability. The configured working calibrator, after calibrating the detection system at the end of the shelf life, has a linear correlation coefficient r2≥0.990 within the test range of 0.5ng / mL-300ng / mL. The CK-MB calibrator can be widely used in various biochemical analyzers and luminometers, has a wider versatility, and solves the problem of unstable storage of existing creatine kinase isoenzyme freeze-dried calibrators. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0016] Figure 1 It is a schematic diagram of clinical test results of a stable creatine kinase isoenzyme freeze-dried product provided by the present invention.

[0017] Figure 2 The present invention provides a flow chart of a method for preparing a stable creatine kinase isoenzyme freeze-dried product. DETAILED DESCRIPTION

[0018] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, and should not be construed as limiting the present invention.

[0019] In a first aspect, the present invention provides a stable creatine kinase isoenzyme freeze-dried product, comprising the following components: 20-100 mmol / L of a buffer solution with a pH of 7.4±0.2, 0.05-0.2% of a preservative, 2-20% of a protein stabilizer, 0.2%-1% of a metal ion stabilizer, 0.01%-0.03% of an inhibitor, 0.1%-1% of an amino acid, 5%-15% of a sugar alcohol stabilizer, 10 mL / L of glycerol, 0.05%-0.2% of a surfactant, and 0-300 ng / mL of a creatine kinase isoenzyme antigen.

[0020] In an embodiment of the present invention, the buffer comprises any one of a MES buffer and a mixed buffer of MES and Tris, the preservative comprises one or more of Proclin 300, NaN3 and gentamicin, the metal ion stabilizer is NaCl, the protein stabilizer comprises one or more of human serum albumin, bovine serum albumin, casein and animal serum, the amino acid comprises any one of glycine and L-arginine, the inhibitor is phenylmethylsulfonyl fluoride, the sugar alcohol stabilizer comprises any two or more of trehalose, sucrose, sorbitol and dextran 500, the surfactant comprises one or two of Triton 100 and Tween 20, the enzyme isozyme antigen concentration is 0 ng / mL, 2 ng / mL, 10 ng / mL, 30 ng / mL, 60 ng / mL and 300 ng / mL, and the creatine kinase isozyme lyophilized product is densely stored at 2°C to 8°C. It can be stored under sealed conditions for 15 months; the shelf life is 8h when placed in the supporting instrument in the open state. It has good detection performance for the determination of creatine kinase isoenzyme (CK-MB). After reconstitution, it can be stored at -20℃ for 1 month and at -20℃ for 2 months. It has stable performance, low cost, accurate test results, good repeatability, and the configured working calibrator, after calibrating the detection system at the end of the validity period, has a linear correlation coefficient of r2≥0.990 within the test range of 0.5ng / mL-300ng / mL under room temperature conditions of 18℃~28℃. The CK-MB calibrator can be widely used in various biochemical analyzers and luminometers, has a wider versatility, and solves the problem of unstable storage of existing creatine kinase isoenzyme freeze-dried calibrators.

[0021] See also Figure 1-Figure 2 In a second aspect, the present invention further provides a method for preparing a stable lyophilized product of creatine kinase isoenzyme, which is used to prepare the stable lyophilized product of creatine kinase isoenzyme as described in the first aspect, and is characterized in that it comprises the following steps:

[0022] S1: Take a beaker and fill it with a pre-prepared volume of purified water, and add creatine kinase isoenzyme antigen and metal ion stabilizer to the beaker in sequence, stir and mix, and adjust the pH to obtain a primary mixed solution;

[0023] In the embodiment of the present invention, at a temperature of 25±3° C., accurately weighed buffer and metal ion stabilizer are dissolved one by one in a pre-prepared volume of purified water, and the pH value of the solution is adjusted to within the range of 7.4±0.5 with 1 mol / L sodium hydroxide solution to obtain a primary mixed solution.

[0024] S2. adding a sugar alcohol stabilizer, an amino acid and a protein stabilizer to the primary mixed solution in sequence, stirring and dissolving, to obtain a secondary mixed solution;

[0025] In the embodiment of the present invention, about 30% of the sugar alcohol stabilizer and the protein stabilizer dissolved in 60° C. purified water are added to the primary mixed solution, and the mixture is stirred and dissolved to obtain a secondary mixed solution.

[0026] S3: adding a sugar alcohol stabilizer, glycerol, an inhibitor, a surfactant and a preservative to the secondary mixed solution in sequence, stirring and dissolving, fixing the volume, filtering with a filter membrane, and obtaining a lyophilized product of creatine kinase isoenzyme.

[0027] In an embodiment of the present invention, glycerol, surfactant and preservative are added to the secondary mixed solution, and then purified water is added to dilute the total amount of the reagent, the volume is fixed, and after standing overnight, it is filtered with a 0.2 μm pore size filter membrane to obtain a secondary mixed solution.

[0028] In order to better understand the present technical solution, the following embodiments are provided for further explanation:

[0029] Embodiment 1:

[0030] CK-MB calibrator includes the following components: MES buffer 100mmol / L, pH7.4, Prolin 3000.1%, bovine serum albumin 30g / L, NaCl 0.9%, trehalose 40g / L, dextran 50020g / L, glycerol 10mL / L and Triton 1000.1%.

[0031] A method for preparing a stable lyophilized product of creatine kinase isoenzyme:

[0032] At 25±3℃, dissolve accurately weighed MES buffer 100mmol / L and NaCl 0.9% one by one with about 60% of the prepared amount of purified water, adjust the solution pH value within the range of 7.4±0.5 with 1mol / L sodium hydroxide solution, add dextran 500 dissolved with about 30% of the prepared amount of 60℃ purified water, bovine serum albumin 30g / L, trehalose 40g / L, glycerol 10mL / L, Triton 1000.1%, Prolin 3000.1%, then dilute with purified water to the total prepared amount of the reagent, let it stand overnight and filter with a 0.2μm pore size filter membrane.

[0033] Table 1-1 Long-term stability monitoring of Example 1

[0034] Calibrator concentration (ng / mL) 1 month 2 months 4 months 8 months 12 months 15 months 16 months 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.99 2.05 1.99 1.99 1.91 1.84 1.94 1.89 9.85 9.38 9.47 10.23 9.96 9.42 9.52 9.31 30.57 29.78 30.87 30.1 29.59 29.65 29.11 28.78 59.86 57.59 59.34 62.1 60.66 56.45 58.7 56.74 299.14 300.78 285.11 293.61 299.35 307.17 283.85 276.49

[0035] Table 1-2 Stability in Example 1

[0036]

[0037] Table 1-3 Real-time stability in Example 1

[0038]

[0039] Table 1-4 Reconstitution 2-8℃ Storage Stability and Deviation in Example 1

[0040]

[0041]

[0042] Table 1-5 Freeze-thaw stability in Example 1

[0043] Concentration (ng / mL) Day 0 7 days 21 days 42 days 56 days 62 days 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.99 1.96 2.05 1.92 2.00 1.94 1.88 9.85 10.21 9.67 9.92 9.96 9.45 9.27 30.57 30.45 30.97 29.37 28.61 28.91 28.22 59.86 60.67 60.11 61.35 59.97 55.74 60.87 299.14 295.15 309.32 310.91 292.76 292.98 302.61 deviation\% Day 0 7 days 14 days 21 days 28 days 31 days 1.99 -1.51％ 3.02％ -3.52％ 0.50％ -2.51％ -5.53％ 9.85 3.65％ -1.83％ 0.71％ 1.12％ -4.06％ -5.89％ 30.57 -0.39％ 1.31％ -3.93％ -6.41％ -5.43％ -7.69％ 59.86 1.35％ 0.42％ 2.49％ 0.18％ -6.88％ 1.69％ 299.14 -1.33％ 3.40％ 3.93％ -2.13％ -2.06％ 1.16％

[0044] Embodiment 2:

[0045] The CK-MB calibrator includes the following components: MES buffer 50mmol / L, pH 7.4, NaN30.05%, bovine serum albumin 30g / L, NaCl 0.9%, sucrose 20g / L, sorbitol 30g / L, trehalose 20g / L, glycerol 10mL / L and Triton 1000.1%.

[0046] A method for preparing a stable lyophilized product of creatine kinase isoenzyme:

[0047] At 25±3℃, dissolve accurately weighed MES buffer 50mmol / L, NaCl 0.9%, adjust the solution pH to 7.4±0.5 with 1mol / L sodium hydroxide solution, sucrose 20g / L, sorbitol 30g / L, trehalose 20g / L, bovine serum albumin 30g / L, NaN30.05%, glycerol 10mL / L respectively with about 80% of the prepared amount of purified water, then dilute with purified water to the total prepared amount of the reagent, let stand overnight and filter with a 0.2μm pore size filter membrane.

[0048] Table 2-1 Long-term stability monitoring of Example 1

[0049] Calibrator concentration (ng / mL) 1 month 2 months 4 months 8 months 12 months 15 months 16 months 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.99 1.95 2.02 2 1.91 1.96 1.86 1.9 9.85 10.17 9.44 9.6 9.26 9.7 9.59 9.09 30.57 30.9 31.75 31.19 29.34 31.92 29.68 28.92 59.86 59.8 58.61 58.07 55.25 60.54 57.36 55.08 299.14 307.94 313.54 305.01 284.77 304.6 279.77 277.56

[0050] Table 2-2 Stability in Example 1

[0051]

[0052]

[0053] Table 2-3 Real-time stability in Example 1

[0054]

[0055] Table 2-4 Reconstitution 2-8℃ Storage Stability and Deviation in Example 1

[0056] Concentration (ng / mL) Day 0 7 days 14 days 21 days 28 days 31 days 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.99 1.99 1.95 1.95 1.89 1.96 1.86 9.85 9.81 9.88 9.68 9.87 9.74 9.71 30.57 30.36 30.60 31.74 29.46 31.00 29.45 59.86 60.47 61.58 56.93 61.87 56.35 56.75 299.14 302.34 286.32 290.47 291.27 300.58 288.64 deviation\% Day 0 7 days 14 days 21 days 28 days 31 days 1.99 0.00％ -2.01％ -2.01％ -5.03％ -1.51％ -6.53％ 9.85 -0.41％ 0.30％ -1.73％ 0.20％ -1.12％ -1.42％ 30.57 -0.69％ 0.10％ 3.83％ -3.63％ 1.41％ -3.66％ 59.86 1.02％ 2.87％ -4.89％ 3.36％ -5.86％ -5.20％ 299.14 1.07％ -4.29％ -2.90％ -2.63％ 0.48％ -3.51％

[0057] Table 2-5 Freeze-thaw stability in Example 1

[0058] Concentration (ng / mL) Day 0 7 days 21 days 42 days 56 days 62 days 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.99 1.98 2.03 1.97 1.94 1.84 1.87 9.85 10.06 10.09 9.62 10.24 9.48 9.18 30.57 31.28 31.98 30.68 30.52 31.66 29.42 59.86 60.62 62.43 62.50 60.22 55.58 59.21 299.14 299.67 292.75 306.87 279.40 285.54 280.56 deviation\% Day 0 7 days 14 days 21 days 28 days 31 days 1.99 -0.50％ 2.01％ -1.01％ -2.51％ -7.54％ -6.03％ 9.85 2.13％ 2.44％ -2.34％ 3.96％ -3.76％ -6.80％ 30.57 2.32％ 4.61％ 0.36％ -0.16％ 3.57％ -3.76％ 59.86 1.27％ 4.29％ 4.41％ 0.60％ -7.15％ -1.09％ 299.14 0.18％ -2.14％ 2.58％ -6.60％ -4.55％ -6.21％

[0059] Embodiment 3:

[0060] CK-MB calibrator includes the following components: MES buffer 100mmol / L, pH7.4, Prolin 3000.1%, bovine serum albumin 35g / L, NaCl 0.9%, trehalose 40g / L, dextran 50020g / L, glycerol 10mL / L and Triton 1000.1%

[0061] A method for preparing a stable lyophilized product of creatine kinase isoenzyme:

[0062] At 25±3℃, dissolve accurately weighed MES 50mmol / L and Tris 20mmol / L, NaCl 0.9% one by one with about 60% of the prepared amount of purified water, adjust the solution pH value to 7.4±0.5 with 1mol / L sodium hydroxide solution, add dextran 500 dissolved with about 30% of the prepared amount of 60℃ purified water, bovine serum albumin 35g / L, trehalose 40g / L, glycerol 10mL / L, Prolin 3000.1%, Triton 1000.1%, then dilute with purified water to the total prepared amount of the reagent, let it stand overnight and filter with a 0.2μm pore size filter membrane.

[0063] Table 3-1 Long-term stability monitoring of Example 1

[0064] Calibrator concentration (ng / mL) 1 month 2 months 4 months 8 months 12 months 15 months 16 months 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.99 1.95 2.02 2.00 1.91 1.96 1.86 1.90 9.85 10.17 9.44 9.60 9.26 9.70 9.59 9.09 30.57 30.90 31.75 31.19 29.34 31.92 29.68 28.92 59.86 59.80 58.61 58.07 55.25 60.54 57.36 55.08 299.14 307.94 313.54 305.01 284.77 304.60 279.77 277.56

[0065] Table 3-2 Stability in Example 1

[0066]

[0067] Table 3-3 Real-time stability in Example 1

[0068]

[0069] Table 3-4 Reconstitution 2-8℃ Storage Stability and Deviation in Example 1

[0070]

[0071]

[0072] Table 3-5 Freeze-thaw stability in Example 1

[0073] Concentration (ng / mL) Day 0 7 days 21 days 42 days 56 days 62 days 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.99 1.98 2.03 1.97 1.94 1.84 1.87 9.85 10.06 10.09 9.62 10.24 9.48 9.18 30.57 31.28 31.98 30.68 30.52 31.66 29.42 59.86 60.62 62.43 62.50 60.22 55.58 59.21 299.14 299.67 292.75 306.87 279.40 285.54 280.56 deviation\% Day 0 7 days 14 days 21 days 28 days 31 days 1.99 -0.50％ 2.01％ -1.01％ -2.51％ -7.54％ -6.03％ 9.85 2.13％ 2.44％ -2.34％ 3.96％ -3.76％ -6.80％ 30.57 2.32％ 4.61％ 0.36％ -0.16％ 3.57％ -3.76％ 59.86 1.27％ 4.29％ 4.41％ 0.60％ -7.15％ -1.09％ 299.14 0.18％ -2.14％ 2.58％ -6.60％ -4.55％ -6.21％

[0074] Freeze drying process

[0075]

[0076] After freeze-drying the samples of Examples 1, 2, and 3, the water content should be within the range of 3% ± 1%. Too much or too little water is not conducive to the long-term storage of the freeze-dried product.

[0077] Example 4

[0078] CK-MB Assay Kit Calibrator Formula

[0079] Including MES buffer 100mmol / L, pH7.4, Prolin 3000.1%, bovine serum albumin 30g / L, NaCl0.9%, trehalose 40g / L, dextran 50020g / L, glycerol 10mL / L and Triton 1000.1%

[0080] Clinical relevance verification: The working calibrator was prepared using the implementation case formula, and compared with the CK-MB assay kit of a company approved by the State Drug Administration on the market at the end of the validity period. The same clinical samples were tested on the Roche luminometer E411 and the company's luminometer IA260 at the same time. 76 samples were tested and the correlation analysis of the two groups of test results was performed. The results showed that the linear correlation coefficient R2 of the two groups of data was greater than 0.9750, indicating that this reagent has a good correlation with similar detection reagents in clinical test results.

[0081] The above disclosure is only a preferred embodiment of a stable creatine kinase isoenzyme freeze-dried product and a preparation method of the present invention. Of course, this cannot be used to limit the scope of the present invention. A person skilled in the art can understand that all or part of the processes of the above embodiments and equivalent changes made according to the claims of the present invention still fall within the scope of the invention.

Claims

1. A stable lyophilized product of creatine kinase isoenzyme, characterized in that: The invention comprises the following components: 20-100 mmol / L of buffer solution with a pH value of 7.4±0.2, 0.05-0.2% of preservative, 2-20% of protein stabilizer, 0.2%-1% of metal ion stabilizer, 0.01%-0.03% of inhibitor, 0.1%-1% of amino acid, 5%-15% of sugar alcohol stabilizer, 10 mL / L of glycerol, 0.05%-0.2% of surfactant and 0-300 ng / mL of creatine kinase isoenzyme antigen.

2. The stable creatine kinase isozyme freeze-dried product as claimed in claim 1, characterized in that ； The buffer comprises any one of a MES buffer and a mixed buffer of MES and Tris, and the preservative comprises one or more of Proclin 300, NaN3 and gentamicin.

3. The stable creatine kinase isozyme freeze-dried product as claimed in claim 1, characterized in that ； The metal ion stabilizer is NaCl, the protein stabilizer includes one or more of human serum albumin, bovine serum albumin, casein and animal serum, the amino acid includes any one of glycine and L-arginine, the inhibitor is phenylmethylsulfonyl fluoride, and the sugar alcohol stabilizer includes any two or more of trehalose, sucrose, sorbitol and dextran 500.

4. The stable creatine kinase isozyme freeze-dried product as claimed in claim 1, characterized in that ； The surfactant includes one or both of Triton 100 and Tween 20, and the concentration of the enzyme isozyme antigen is 0 ng / mL, 2 ng / mL, 10 ng / mL, 30 ng / mL, 60 ng / mL and 300 ng / mL.

5. A method for preparing a stable lyophilized product of creatine kinase isoenzyme, for preparing the stable lyophilized product of creatine kinase isoenzyme as claimed in any one of claims 1 to 4, characterized in that: The following steps are involved: Take a beaker and fill it with a pre-prepared volume of purified water, and add creatine kinase isoenzyme antigen and metal ion stabilizer to the beaker in sequence, stir and mix, adjust the pH, and obtain a primary mixed solution; Adding a sugar alcohol stabilizer, an amino acid and a protein stabilizer to the primary mixed solution in sequence, stirring and dissolving, to obtain a secondary mixed solution; Add sugar alcohol stabilizer, glycerol, inhibitor, surfactant and preservative to the secondary mixed solution in sequence, stir and dissolve, adjust the volume, filter with a filter membrane, and obtain creatine kinase isoenzyme freeze-dried product.

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