Composition, protein control substrate solution containing the same, CHI3L1 control, CHI3L1 detection kit and detection method

Abstract

The application relates to the technical field of biological medicine, in particular to a composition, a protein control matrix liquid containing the same, CHI3L1 control liquid, a CHI3L1 detection kit and a detection method. The composition comprises glycine and trehalose in a weight ratio of (19-21):(8.5-11.5). Compared with the traditional technology, the application has the following beneficial effects: the glycine and the trehalose are matched in a specific ratio to form a specific composition, the CHI3L1 control liquid is prepared by using the protein control matrix liquid added with the composition, the performance is stable, in particular, the control liquid does not need to be freeze-dried (i.e. stored in a liquid state), and after being opened, the control liquid can still remain stable, and the current detection requirements can be met.

Description

Technical Field

[0001] This application relates to the field of biomedical technology, and in particular to a composition, a protein control matrix solution containing the composition, a CHI3L1 quality control product, a CHI3L1 detection kit, and a detection method. Background Technology

[0002] Currently, the main methods for detecting liver fibrosis in clinical practice include invasive and non-invasive examinations.

[0003] Invasive examinations primarily refer to liver biopsy, the gold standard for diagnosing liver fibrosis, which can assess the degree of inflammatory response in liver tissue and the stage of liver fibrosis. However, in clinical practice, because liver biopsy is an invasive procedure, it can cause complications such as bleeding and bile leakage, making it difficult for most patients to tolerate. Furthermore, it requires physicians with extensive experience in liver biopsy and a relevant department with high-level pathology expertise. These factors limit the widespread clinical application of liver biopsy.

[0004] Non-invasive examinations, including imaging studies such as ultrasound, CT, and MRI, are not very valuable for diagnosing early-stage liver fibrosis because they lack specific signs of the condition. Furthermore, clinically relevant serum diagnostic markers for liver fibrosis mainly include laminin (LN), hyaluronic acid (HA), type IV collagen (CIV), and type III collagen (PⅢNP). However, these levels are easily affected by hepatocellular necrosis and inflammatory damage, and may also be elevated in other diseases, thus failing to accurately identify liver fibrosis.

[0005] Chitinase 3-like 1 (CHI3L1), also known as HC-gp39, YKL-40, and mammary gland degeneration protein 39, was initially discovered in the supernatant of MG63 cell line culture. Its polypeptide chain has tyrosine (Y), lysine (K), and leucine (L) at its amino terminals, with a relative molecular mass of approximately 40 kDa, hence the initial name YKL-40, later changed to CHI3L1. The gene and protein sequences of CHI3L1 were elucidated in 1993. Its encoding gene is located in the lq31-q32 region of chromosome 1, and is a 7948-base DNA fragment containing 10 exons. Studies have shown that CHI3L1 is expressed in organs such as the heart and brain, but it is highly expressed in the liver. The expression level in the human heart is about 15 times higher than that in the kidneys and more than 200 times higher than that in the heart, which is 15 to 227 times higher than that in other tissues. Therefore, its application value in liver diseases has received increasing attention and it is considered a useful biomarker of liver fibrosis in different contexts.

[0006] Currently, there are relatively few detection methods and products for chitosan polysaccharide enzyme 3-like protein 1 on the market, with serological testing being the primary method. Products for detecting antigens and antibodies using methods such as colloidal gold assay and magnetic particle chemiluminescence assay are available, with magnetic particle chemiluminescence assay representing the main development trend.

[0007] In recent years, there have been very few quality control products of chitosan polysaccharide enzyme 3-like protein 1, which cannot better meet market demand.

[0008] Therefore, this application is hereby submitted. Summary of the Invention

[0009] Based on this, one or more embodiments of this application provide a composition, a protein control matrix solution containing the same, a CHI3L1 quality control sample, a CHI3L1 detection kit, and a detection method. The technical solutions include the following:

[0010] One or more embodiments of this application provide a composition comprising glycine and trehalose in a weight ratio of (19-21):(8.5-11.5).

[0011] In some embodiments of this application, the composition comprises glycine and trehalose in a weight ratio of (19.5–20.5):(9.5–10.5).

[0012] One or more embodiments of this application provide a protein control matrix solution, the protein matrix solution comprising the composition described above.

[0013] In some embodiments of this application, the protein control matrix solution includes glycine, trehalose, and fetal bovine serum;

[0014] Optionally, the weights of glycine and trehalose corresponding to each 1L of the fetal bovine serum are 19g-21g and 8.5g-11.5g, respectively.

[0015] Optionally, the weights of glycine and trehalose corresponding to each 1L of the fetal bovine serum are 19.5g to 20.5g and 9.5g to 10.5g, respectively.

[0016] In some embodiments of this application, the protein control matrix solution further includes a preservative;

[0017] Optionally, the preservative includes PC300;

[0018] Optionally, the preservative content per 1L of fetal bovine serum is 0.3g to 0.5g.

[0019] One or more embodiments of this application provide a CHI3L1 quality control sample, the CHI3L1 quality control sample comprising:

[0020] The composition or the protein control matrix solution; and,

[0021] CHI3L1.

[0022] In some embodiments of this application, the CHI3L1 quality control product is a combination product, which includes multiple quality control products and the concentration of CHI3L1 in the multiple quality control products is different.

[0023] In some embodiments of this application, the combined product includes two quality control samples; optionally, the combined product includes:

[0024] The first concentration control sample, wherein the concentration of CHI3L1 is 30 ng / mL to 50 ng / mL; and,

[0025] The second concentration control sample contains CHI3L1 at a concentration of 150 ng / mL to 250 ng / mL.

[0026] One or more embodiments of this application provide a CHI3L1 detection kit, wherein the CHI3L1 detection kit includes the CHI3L1 quality control sample.

[0027] One or more embodiments of this application provide a method for detecting CHI3L1 in a sample, wherein the detection method uses the CHI3L1 quality control material or the CHI3L1 detection kit in the process of detecting CHI3L1 in the sample.

[0028] Compared with traditional technologies, this application has the following advantages:

[0029] This application combines glycine and trehalose in a specific ratio to form a specific composition. The CHI3L1 quality control sample is prepared using a protein control matrix solution containing this composition. The quality control sample is stable, and in particular, it does not require lyophilization (i.e., it is stored in liquid form) and remains stable even after opening and re-storage, which can meet current testing requirements. Detailed Implementation

[0030] The present application will be further described in detail below with reference to the embodiments and examples. It should be understood that these embodiments and examples are for illustrative purposes only and are not intended to limit the scope of the present application. The purpose of providing these embodiments and examples is to enable a more thorough and comprehensive understanding of the disclosure of the present application. It should also be understood that the present application can be implemented in many different forms and is not limited to the embodiments and examples described herein. Those skilled in the art can make various modifications or alterations without departing from the spirit of the present application, and the equivalent forms obtained also fall within the protection scope of the present application. Furthermore, numerous specific details are set forth in the following description to provide a more complete understanding of the present application. It should be understood that the present application can be implemented without one or more of these details.

[0031] 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 application belongs. The terminology used herein in the specification of this application is for descriptive purposes only and is not intended to be limiting of the application.

[0032] the term

[0033] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0034] The terms "and / or," "or / and," and "and / or" as used herein include any one of two or more of the related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that, in this application, the technical solution undoubtedly includes technical solutions connected by "logical AND," and also undoubtedly includes technical solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0035] In this application, the terms "multiple", "various", "multiple times", "multi-dimensional", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0036] The terms “combinations of,” “any combination of,” and “any combination of” used in this article include all suitable combinations of any two or more of the listed items.

[0037] In this document, the term "suitable" as used in phrases such as "suitable combination," "suitable method," and "any suitable method" refers to the ability to implement the technical solution of this application, solve the technical problem of this application, and achieve the expected technical effect of this application.

[0038] In this document, terms such as “preferred,” “better,” “more suitable,” and “ideal” are merely used to describe implementation methods or examples that achieve better results, and should be understood not to limit the scope of protection of this application.

[0039] In this application, terms such as "further," "even further," and "particularly" are used to describe purposes and indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0040] In this application, "optionally," "optionally," and "optional" mean that something is optional, that is, it means that it is selected from either "with" or "without." If there are multiple "optional" entries in a technical solution, unless otherwise specified, and there are no contradictions or mutual constraints, each "optional" entry shall be independent.

[0041] In this application, the terms "first aspect," "second aspect," "third aspect," "fourth aspect," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first," "second," "third," "fourth," etc., serve only as a non-exhaustive enumeration and should be understood not to constitute a closed limitation on quantity.

[0042] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0043] In this application, numerical intervals (i.e., numerical ranges) are involved. Unless otherwise specified, the selected numerical distributions within the aforementioned numerical intervals are considered continuous and include the two endpoints (i.e., the minimum and maximum values) of the numerical range, as well as every value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints. In this document, this is equivalent to directly listing every integer. For example, if t is an integer selected from 1 to 10, it means that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. Furthermore, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0044] Unless otherwise specified, the temperature parameters in this application are permitted to be either constant-temperature treatment or variations within a certain temperature range. It should be understood that the constant-temperature treatment allows temperature fluctuations within the precision range of the instrument control, such as ±5℃, ±4℃, ±3℃, ±2℃, or ±1℃.

[0045] In this application, % (w / w) and wt% both represent weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass-volume percentage.

[0046] All references to this application are incorporated herein by reference as if each document were individually incorporated herein by reference. Unless they conflict with the purpose and / or technical solution of this application, all cited references are incorporated herein by reference in their entirety and for all purposes. When references are cited in this application, the definitions of relevant technical features, terms, nouns, phrases, etc., are also incorporated herein by reference. Examples and preferred embodiments of the cited technical features may also be incorporated herein by reference, but only to the extent that they enable the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, this application shall prevail or modifications shall be made adaptably to the description in this application.

[0047] A first aspect of this application provides a composition comprising glycine and trehalose in a weight ratio of (19-21):(8.5-11.5).

[0048] This application combines glycine and trehalose in a specific ratio to form a specific composition. The CHI3L1 quality control sample is prepared using a protein control matrix solution containing this composition. The quality control sample is stable, and in particular, it does not require lyophilization (i.e., it is stored in liquid form) and remains stable even after opening and re-storage, which can meet current testing requirements.

[0049] In this application, the weight ratio of glycine to trehalose in the composition is, for example, 19:8.5, 19.5:8.5, 20:8.5, 20.5:8.5, 21:8.5, 19:9, 19.5:9, 20:9, 20.5:9, 21:9, 19:10.5, 19.5:10.5, 20:10.5, 20.5:10.5, 21:10.5, 19:11, 19.5:11, 20:11, 20.5:11, 21:11, 19:11.5, 19.5:11.5, 20:11.5, 20.5:11.5, 21:11.5.

[0050] In some examples of this application, the composition comprises glycine and trehalose in a weight ratio of (19.5–20.5):(9.5–10.5).

[0051] A second aspect of this application provides a protein control matrix solution, the protein matrix solution comprising the aforementioned composition.

[0052] In some examples of this application, the protein control matrix solution includes glycine, trehalose, and fetal bovine serum.

[0053] First, the CHI3L1 quality control product of this application uses animal serum, which greatly reduces costs while maintaining high accuracy, and can be produced in batches to meet the needs of clinical clients.

[0054] Secondly, the CHI3L1 quality control product of this application can be a liquid, which facilitates subsequent testing and makes it easier for hospitals and testing institutions to verify and evaluate it. Furthermore, the shelf life of the CHI3L1 quality control product after opening is significantly longer than that of currently available freeze-dried quality control products on the market, while still maintaining excellent performance to meet market demands.

[0055] In some examples of this application, the weights of glycine and trehalose corresponding to each 1L of fetal bovine serum are 19g to 21g (e.g., 19, 19.5, 20, 20.5, 21g) and 8.5g to 11.5g (e.g., 8.5, 9, 9.5, 10, 10.5, 11, 11.5g), respectively.

[0056] In some examples of this application, the weights of glycine and trehalose corresponding to each 1L of the fetal bovine serum are 19.5g to 20.5g and 9.5g to 10.5g, respectively.

[0057] In some examples of this application, the protein control matrix solution also includes a preservative. This application does not specifically limit the type of preservative, but includes, but is not limited to, PC300. This application does not specifically limit the amount of preservative used; an appropriate amount of preservative can be added to the protein control matrix solution, including, but not limited to, 0.3g to 0.5g (e.g., 0.3, 0.35, 0.4, 0.45, 0.5g) of preservative per 1L of fetal bovine serum.

[0058] A third aspect of this application provides a CHI3L1 quality control sample, the CHI3L1 quality control sample comprising:

[0059] The composition or the protein control matrix solution; and,

[0060] CHI3L1.

[0061] In some examples of this application, the CHI3L1 quality control sample is a combination product, which includes multiple quality control samples with different concentrations of CHI3L1 in the multiple quality control samples. This application does not specifically limit the quality control samples, for example, 2, 3, 4, 5, 6, and 7.

[0062] In some examples of this application, the combined product includes two quality control samples, wherein the quality control sample with a relatively high concentration of CHI3L1 can be referred to as a high-concentration quality control sample, and the quality control sample with a relatively low concentration can be referred to as a low-concentration quality control sample.

[0063] In some examples of this application, the combined product includes:

[0064] The first concentration quality control sample, wherein the concentration of CHI3L1 is 30 ng / mL to 50 ng / mL (e.g., 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50 ng / mL); and,

[0065] The second concentration quality control sample contains CHI3L1 at a concentration of 150 ng / mL to 250 ng / mL (e.g., 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250 ng / mL).

[0066] A fourth aspect of this application provides a CHI3L1 detection kit, the CHI3L1 detection kit including the aforementioned CHI3L1 quality control sample.

[0067] A fifth aspect of this application provides a method for detecting CHI3L1 in a sample, wherein the detection method uses the CHI3L1 quality control material or the CHI3L1 detection kit during the detection of CHI3L1 in the sample.

[0068] The embodiments of this application will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this application, or follow experimental manuals or conventional conditions in the art, or follow the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0069] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. For temperature and time parameters, acceptable deviations due to instrument testing accuracy or operational precision are permissible.

[0070] The following embodiments are further illustrations of the present invention, but not limitations thereof. Unless otherwise specified, the equipment and reagents used in this invention are commercially available products conventional in this technical field.

[0071] Example 1: Adding trehalose and glycine to the quality control matrix solution effectively improved the stability of the quality control products for this project.

[0072] The quality control matrix solution is divided into two portions, numbered 1# and 2# respectively. The matrix solution of 1# does not contain any excipients and consists of fetal bovine serum and PC300. The amount of PC300 used per 1L of fetal bovine serum is 0.5g. The matrix solution of 2# contains a certain proportion of trehalose and glycine and consists of fetal bovine serum, glycine, trehalose and PC300. The amount of glycine, trehalose and PC300 used per 1L of fetal bovine serum is 20g, 10g and 0.5g respectively. This is designated as 2#'.

[0073] Divide #1 into two portions of quality control matrix solution, and label them #3 and #4 respectively; divide #2 into two portions of quality control matrix solution, and label them #5 and #6 respectively;

[0074] Add an equal and low amount of CHI3L1 antigen to the above 3# and 5# to form CHI3L1 quality control products 3# and 5#;

[0075] Add an equal and high amount of CHI3L1 antigen to the above 4# and 6# to form CHI3L1 quality control products 4# and 6#.

[0076] Dispense the samples into 1 mL / borosilicate transparent glass bottles. Three replicates were set up for each test condition, and the average value was taken for subsequent tests.

[0077] The concentrations of CHI3L1 antigen were measured after 10 days of accelerated heating at 37℃, 10 days of lyophilization and reconstitution at 2℃~8℃, 10 days at -20℃, 10 days at -80℃, and 3 days of storage at 25℃, and the relative change rate was calculated.

[0078] The concentration of CHI3L1 antigen in the CHI3L1 quality control sample was determined using a chitosan polysaccharide enzyme 3-like protein 1 assay kit (chemiluminescence method) and an AHILONG chemiluminescence analyzer. The test results and the corresponding calculated relative change rates are shown in Table 1.

[0079] Table 1 (CHI3L1 antigen concentration unit: ng / mL)

[0080] Quality control solution Initial concentration of CHI3L1 antigen 37℃ - 10 days 25℃-3d 2℃～8℃-10d -20℃-10d -80℃-10d 3# 40.81 36.22 41.24 41.22 42.15 41.91 relative deviation / -11.25％ 1.05％ 1.00％ 3.28％ 2.70％ 5# 39.34 40.06 39.23 39.73 39.30 39.81 relative deviation / 1.83％ -0.28％ 0.99％ -0.10％ 1.19％ 4# 221.41 188.26 208.23 218.56 221.78 220.55 relative deviation / -14.97％ -5.95％ -1.29％ 0.17％ -0.39％ 6# 205.7 194.42 204.07 209.17 214.08 213.5 relative deviation / -5.48％ -0.79％ 1.69％ 4.07％ 3.79％

[0081] Table 1 shows that for the CHI3L1 quality control samples, comparing the results of samples #3 and #5, and #4 and #6, the quality control test results for #5 and #6 indicate that adding glycine and trehalose to the matrix solution effectively improves the thermal stability of the quality control samples compared to using the matrix solution directly. Specifically, after being treated at 37℃ for 10 days, the relative deviation of quality control sample #3 was -11.25%, and that of quality control sample #5 was 1.83%; the relative deviation of quality control sample #4 was -14.97%, and that of quality control sample #6 was -5.48%. This demonstrates that adding a certain proportion of trehalose and glycine to the matrix solution of the quality control samples effectively improves their thermal stability.

[0082] Example 2: Preparation process of chitosan polysaccharide enzyme 3-like protein 1 quality control material

[0083] A method for preparing a quality control product of chitosan polysaccharide enzyme 3-like protein 1 includes the following steps:

[0084] (1) Raw material processing and concentration confirmation: The concentration of CHI3L1 antigen raw material was determined using a magnetic microparticle chemiluminescence detection kit.

[0085] (2) Preparation of Quality Control Matrix Solution: The quality control matrix solution was prepared in a Class 100,000 production workshop. The preparation process is as follows: Taking 1L of quality control matrix solution as an example, the amounts of each component (2wt% glycine + 1wt% trehalose + 0.05wt% PC300) were calculated and weighed. Then, 1L of fetal bovine serum solution was prepared, and the weighed PC-300, glycine, and trehalose were added to the fetal bovine serum solution in sequence. The mixture was stirred and mixed for 15-20 minutes until the solution was clear and transparent.

[0086] (3) Preparation of quality control samples: Add an appropriate amount of the above antigens to the quality control sample matrix solution, detect the level of each antigen using magnetic microparticle chemiluminescence method, and label it to obtain the following concentration combination of quality control samples;

[0087] The quality control at level 1 contains: 40 ng / mL chitosanase 3-like protein 1 (CHI3L1);

[0088] The quality control sample at level 2 contains: 200 ng / mL chitosanase 3-like protein 1 (CHI3L1);

[0089] The quality control samples at levels 1 and 2 above are tested separately. If the test results meet the following quality standards, the next step of packaging is carried out. If the test results do not meet the following standards, the concentration value can be adjusted by adding chitosan polysaccharide enzyme 3-like protein 1 antigen until it meets the following quality standards.

[0090] Quality standards: The quality control of level 1 contains chitosanase 3-like protein 1 antigen: chitosanase 3-like protein 1 (CHI3L1) (36-44) ng / mL; the quality control of level 2 contains chitosanase 3-like protein 1 antigen: chitosanase 3-like protein 1 (CHI3L1) (180-220) ng / mL.

[0091] (4) Packaging of quality control products

[0092] Dispense the above quality control samples into selected clean glass bottles at a specification of 2 mL / vial using an electric continuous dispensing apparatus, and then add rubber stoppers.

[0093] Example 3: Stability verification of chitosanase 3-like protein 1 quality control product

[0094] 1. Thermal acceleration stability

[0095] To verify the performance of the above-mentioned chitosan polysaccharide enzyme 3-like protein 1 quality control product in extreme environments such as high temperature, this embodiment placed the quality control product that had been opened and then closed at 37°C for 11 days for heat treatment, and used the unopened quality control product (with a shelf life of 18 months) stored at 2°C to 8°C as a control.

[0096] The chitosan polysaccharide enzyme 3-like protein 1 (CHI3L1) detection kit (chemiluminescence method) was used to determine the biomarker luminescence value and concentration value of the above-mentioned chitosan polysaccharide enzyme 3-like protein 1 quality control sample under various conditions on the chemiluminescence platform. The stability was good, and the relative deviation was within ±10%. The results are shown in the table below.

[0097] Table 2. Accelerated stability results of chitosanase 3-like protein 1 quality control (level 1)

[0098]

[0099] Table 3. Accelerated stability results of chitosanase 3-like protein 1 quality control (level 2)

[0100]

[0101] 2. Opening stability verification of chitosan polysaccharide enzyme 3-like protein 1 quality control product

[0102] To determine the stability of the above-mentioned chitosan polysaccharide enzyme 3-like protein 1 quality control product after opening (stored closed under sterile conditions after opening) under different storage conditions, this embodiment uses the quality control product after opening and storing it at 25°C for 3 days, 2°C~8°C for 3 days, 20 days and 30 days, and -20°C for 3 days, 20 days, 30 days and 90 days, respectively. The quality control product stored at 2°C~8°C (unopened, with a shelf life of 18 months) is used as a control.

[0103] The chitosan polysaccharide enzyme 3-like protein 1 (CHI3L1) detection kit (chemiluminescence method) was used to determine the biomarker luminescence value and concentration value of the above-mentioned chitosan polysaccharide enzyme 3-like protein 1 quality control sample under various conditions on the chemiluminescence platform. The stability was good, and the relative deviation was within ±10%. The results are shown in the table below:

[0104] Table 4. Results of open-pack stability of chitosan polysaccharide enzyme 3-like protein 1 quality control (level 1)

[0105] Processing time result relative deviation Comparison (unopened) 41.24 / Store at 25°C for 3 days after opening. 41.96 1.75％ Store at 2-8℃ for 3 days after opening. 42.01 1.87％ Store at -20℃ for 3 days after opening. 41.84 1.45％ Comparison (unopened) 41.00 / Store at 2-8℃ for 20 days after opening. 40.47 -1.29％ Store at -20℃ for 20 days after opening. 41.37 0.90％ Comparison (unopened) 41.32 / Store at 2-8℃ for 30 days after opening. 40.39 -2.25％ Store at -20℃ for 30 days after opening. 42.29 2.35％ Comparison (unopened) 43.64 / Store at -20℃ for 90 days after opening. 41.52 -4.87％

[0106] Table 5. Results of open-pack stability of chitosan polysaccharide enzyme 3-like protein 1 quality control (level 2)

[0107]

[0108]

[0109] Example 4: Application of Chitosan Polysaccharide Enzyme 3-like Protein 1 Quality Control Product

[0110] (1) Preparation of quality control samples: Remove the quality control samples from the packaging box and allow them to equilibrate to room temperature. Carefully open the bottle stopper to avoid liquid spillage.

[0111] (2) Accuracy testing of quality control samples: Using a chemiluminescence analyzer, the above-mentioned shaken and mixed quality control samples were used as samples, and the CHI3L1 item was tested 3 times. The average value was calculated, and the deviation between the average value and the given target value was calculated to examine the accuracy of the quality control samples. The accuracy results all met the following requirements: the relative deviation was within ±10%; the results are shown in the table below.

[0112] Table 6. Accuracy test data of chitosan polysaccharide enzyme 3-like protein 1 quality control product

[0113]

[0114] Table 6 shows that the deviations of each item in the chitosanase 3 protein 1 quality control level 1 and level 2 are -0.54% and -3.37%, respectively. This indicates that the deviation between the detected value and the calibrated target value of the quality control does not exceed 10%, and the accuracy of the quality control at levels 1 and 2 meets the standard.

[0115] Example 5:

[0116] First, referring to Example 2, prepare the CHI3L1 quality control samples shown in the table below.

[0117] Table 7

[0118]

[0119] Secondly, stability tests were conducted according to Example 3, and the results are shown below:

[0120] Table 8. Accelerated stability results of chitosan polysaccharide enzyme 3-like protein 1 combination 1-4 quality control samples (level 1)

[0121]

[0122] Table 9. Accelerated stability results of chitosan polysaccharide enzyme 3-like protein combination 1-4 quality control samples (level 2)

[0123]

[0124] Table 10. Results of open-pack stability of chitosan polysaccharide enzyme 3-like protein combination 1-4 quality control samples (level 1)

[0125]

[0126]

[0127]

[0128] Table 11. Results of open-pack stability of chitosan polysaccharide enzyme 3-like protein combination 1-4 quality control samples (level 2)

[0129]

[0130]

[0131] Comparative Example 1:

[0132] First, referring to Example 2, a CHI3L1 quality control sample was prepared, wherein...

[0133] Control control combination 1 includes control control 1-low and control control 1-high. The difference between control control 1 and control control 2 in Example 2 is that arginine is used instead of glycine.

[0134] Control control combination 2 includes control control 2-low and control control 2-high. The difference between control control 2 and control control 2 in Example 2 is that sucrose is used instead of trehalose.

[0135] Control control combination 3 includes control control 3-low and control control 3-high. The difference between control control 3-low and control control 3-high and the control control 3-high in Example 2 is that the concentration of glycine per 1L of fetal bovine serum is 21.5g.

[0136] Control group 4 includes control group 4-low and control group 4-high. The difference between control group 4-low and control group 4-high and control group 4-high is that the concentration of trehalose per 1L of fetal bovine serum is 12g.

[0137] Secondly, stability tests were conducted according to Example 3, and the results are shown below:

[0138] Table 12. Accelerated stability results of chitosan polysaccharide enzyme 3-like protein 1 control quality control combination 1-4 (level 1)

[0139]

[0140] Table 13. Accelerated stability results of chitosan polysaccharide enzyme 3-like protein 1 control control combination 1-4 (level 1)

[0141]

[0142]

[0143] Table 14. Results of Opening Stability of Chitosan Polysaccharide Enzyme 3-like Protein 1 Control and Quality Control Combinations 1-4 (Level 1)

[0144]

[0145]

[0146]

[0147] Table 15. Results of Opening Stability of Chitosan Polysaccharide Enzyme 3-like Protein 1 Control and Quality Control Combinations 1-4 (Level 2)

[0148]

[0149]

[0150] In summary, the quality control materials of this application involve important biomarkers for the diagnosis of liver fibrosis, providing more stable, convenient, and lower-cost quality control substances for liver fibrosis detection reagents. The chitosanase 3-like protein 1 quality control material prepared in this application uses a liquid refrigeration method, preparing the chitosanase 3-like protein 1 quality control as a liquid quality control, which greatly facilitates the operation and report output of small and medium-sized hospitals or testing institutions. The matrix solution formula of the quality control material prepared in this application can greatly improve the thermal stability and opening stability of CHI3L1, and even in terms of shelf life, it has a longer shelf life than commercially available lyophilized quality control materials. The quality control materials prepared by this invention have lower costs and better meet the market's demand for commercialized batch production, ensuring clinical application and laboratory testing efficiency.

[0151] The technical features of the above-described embodiments and examples can be combined in any suitable manner. For the sake of brevity, not all possible combinations of the technical features in the above-described embodiments and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0152] The embodiments described above are merely illustrative of several implementation methods of this application, intended to facilitate a detailed understanding of the technical solutions of this application, but should not be construed as limiting the scope of protection of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the scope of protection of this application. Furthermore, it should be understood that after reading the above teachings of this application, those skilled in the art can make various alterations or modifications to this application, and the equivalent forms obtained also fall within the scope of protection of this application. It should also be understood that technical solutions obtained by those skilled in the art based on the technical solutions provided in this application through logical analysis, reasoning, or limited experimentation are all within the scope of protection of the appended claims. Therefore, the scope of protection of this patent application should be determined by the content of the appended claims, and the specification can be used to interpret the content of the claims.

Claims

1. A protein control matrix solution, characterized in that, The protein control matrix solution is composed of glycine, trehalose, fetal bovine serum, and PC300. The weights of glycine and trehalose corresponding to each 1L of the fetal bovine serum are 20g and 10g, respectively. The weight of the PC300 corresponding to each 1L of the fetal bovine serum is 0.5g; The protein control matrix solution is used to prepare CHI3L1 quality control material.

2. A CHI3L1 quality control product, characterized in that, The CHI3L1 quality control products include: The protein control matrix solution according to claim 1; and, CHI3L1.

3. The CHI3L1 quality control product according to claim 2, characterized in that, The CHI3L1 quality control sample is a combination product, which includes multiple quality control samples with different concentrations of CHI3L1 in each sample.

4. The CHI3L1 quality control product according to claim 3, characterized in that, The combined product includes two quality control samples.

5. The CHI3L1 quality control product according to claim 4, characterized in that, The combined product includes: The first concentration control sample, wherein the concentration of CHI3L1 is 30 ng / mL to 50 ng / mL; and, The second concentration control sample contains CHI3L1 at a concentration of 150 ng / mL to 250 ng / mL.

6. A CHI3L1 detection kit, characterized in that, The CHI3L1 test kit includes the CHI3L1 quality control material as described in any one of claims 2 to 5.

7. A method for detecting CHI3L1 in samples not intended for diagnostic purposes, characterized in that, The detection method uses the CHI3L1 quality control material as described in any one of claims 2 to 5 or the CHI3L1 detection kit as described in claim 6 during the detection of CHI3L1 in the sample.

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