Composition, protein control substance matrix liquid containing composition, CHI3L1 quality control substance, CHI3L1 detection kit and detection method

Abstract

The invention relates to the technical field of biological medicines, in particular to a composition, a protein control substance matrix liquid containing the composition, a CHI3L1 quality control substance, a CHI3L1 detection kit and a detection method. The composition comprises glycine and trehalose according to a weight ratio of (19 to 21): (8.5 to 11.5). Compared with the prior art, the invention has the following beneficial effects: the glycine and the trehalose are matched according to a specific ratio to form the specific composition, the CHI3L1 quality control product is prepared by using the protein control product matrix liquid added with the composition, the performance is stable, particularly, the quality control product does not need to be freeze-dried (namely stored in a liquid state) and still keeps stable after being unsealed and stored, and the quality control product has a good application prospect. And the current detection requirements can be met.

Description

Technical Field

[0001] This application relates to the field of biomedical technologies, and particularly relates to a composition, a protein control product matrix solution containing the same, a CHI3L1 quality control product, a CHI3L1 detection kit, and a detection method. Background Art

[0002] Currently, the methods for detecting liver fibrosis clinically mainly include invasive examinations and non-invasive examinations.

[0003] Invasive examinations mainly refer to "liver biopsy", which is the gold standard for diagnosing liver fibrosis and can measure the degree of inflammatory reaction in liver tissue and the staging of liver fibrosis. However, considering the actual clinical situation, since liver biopsy is an invasive examination, it can cause complications such as bleeding and bile fistula, and most patients are difficult to accept it. Moreover, for physicians, it requires physicians with very rich liver biopsy experience and relevant departments with high pathological detection levels. These points above limit the wide application of liver biopsy clinically.

[0004] Non-invasive examinations include imaging examinations, including common B-ultrasound, CT, MRI, etc. Since they have no specific signs for liver fibrosis, their value for diagnosing early liver fibrosis is not high. In addition, the serum diagnostic indicators for liver fibrosis clinically mainly include laminin (LN), hyaluronic acid (HA), type C IV collagen (CIV), and type III procollagen aminoterminal peptide (PIIINP). However, they are easily affected during hepatocyte necrosis and inflammatory injury, and may also increase in other diseases, and cannot accurately distinguish liver fibrosis.

[0005] Chitinase 3-like 1 (CHI3L1), also known as HC-gp39, YKL-40, and breast regression protein 39, was initially discovered in the in vitro culture supernatant of the MG63 cell line. The amino terminus of one of its peptide chains is tyrosine (Y), lysine (K), and leucine (L) respectively, and the relative molecular mass is about 40 kDa. Thus, it was named YKL-40 and later also called CHI3L1. The gene and protein sequences of CHI3L1 were elucidated in 1993. Its encoding gene is located in the chromosomal region 1q31-q32, and it is a DNA fragment with a length of 7948 bases and contains 10 exons. Some studies have shown that CHI3L1 is expressed in organs such as the heart and brain, but is highly expressed in the liver. Its expression level in the human heart is about 15 times higher than that in the kidney, more than 200 times higher than that in the heart, that is, 15 to 227 times that of other tissues. Therefore, its application value in liver diseases has attracted increasing attention, and it is considered a useful biomarker for liver fibrosis in different backgrounds.

[0006] At present, there are few detection methods and detection products for chitinase 3-like protein 1 on the market, mainly serological detection. There are related serological detection products for antigens and antibodies by colloidal gold method, magnetic particle chemiluminescence method, etc., and the magnetic particle chemiluminescence method is the main development trend.

[0007] In recent years, the quality control products for chitinase 3-like protein 1 are extremely scarce and cannot better meet the market demand.

[0008] In view of this, this application is specifically proposed. Summary of the Invention

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

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

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

[0012] One or more embodiments of this application provide a protein quality control product matrix solution, and the protein matrix solution includes the above composition.

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

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

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

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

[0017] Optionally, the preservative includes PC300;

[0018] Optionally, the weight of the preservative corresponding to each 1 L of the fetal bovine serum is 0.3 g-0.5 g.

[0019] One or more embodiments of this application provide a CHI3L1 quality control product, and the CHI3L1 quality control product includes:

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

[0021] CHI3L1.

[0022] In some embodiments of the present application, the CHI3L1 quality control product is a combined product, and the combined product includes a plurality of quality control products with different concentrations of CHI3L1 in the plurality of quality control products.

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

[0024] A first concentration quality control product, wherein the concentration of CHI3L1 therein is 30 ng / mL to 50 ng / mL; and,

[0025] A second concentration quality control product, wherein the concentration of CHI3L1 therein is 150 ng / mL to 250 ng / mL.

[0026] One or more embodiments of the present application provide a CHI3L1 detection kit, and the CHI3L1 detection kit includes the CHI3L1 quality control product described above.

[0027] One or more embodiments of the present application provide a method for detecting CHI3L1 in a sample, and the detection method uses the CHI3L1 quality control product or the CHI3L1 detection kit described above during the process of detecting CHI3L1 in the sample.

[0028] Compared with the traditional technology, the present application has the following beneficial effects:

[0029] The present application combines glycine and trehalose in a specific ratio to form a specific composition, and uses a protein quality control matrix solution added with this composition to prepare a CHI3L1 quality control product, which has stable performance. In particular, the quality control product does not need to be freeze-dried (i.e., stored in liquid state), and still remains stable after being opened and stored again, and can meet the current detection requirements. Detailed implementation manners

[0030] The present application will be further described in detail below in conjunction with embodiments and examples. It should be understood that these embodiments and examples are only used to illustrate the present application and not to limit the scope of the present application. The purpose of providing these embodiments and examples is to make the disclosure of the present application more thoroughly and comprehensively understood. 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 changes or modifications without departing from the connotation of the present application, and the equivalent forms obtained also fall within the protection scope of the present application. In addition, in the following description, a large number of specific details are given to provide a more thorough 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 those of ordinary skill in the technical field to which this application belongs. The terms used in the specification of this application herein are only for the purpose of describing embodiments and examples and are not intended to limit this application.

[0032] The term

[0033] Unless otherwise stated or there is a contradiction, the terms or phrases used herein have the following meanings:

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

[0035] In this application, the terms “multiple”, “diverse”, “multiple times”, “multiple elements”, etc., unless otherwise specified, refer to a quantity greater than 2 or equal to 2. For example, “one or more” means one or greater than or equal to two.

[0036] As used herein, "its combination", "any combination thereof", "any combination mode thereof", etc. include all suitable combination modes of any two or more of the listed items.

[0037] In this application, the "suitable" in "suitable combination mode", "suitable mode", "any suitable mode", etc. is subject to being able to implement the technical solution of this application, solve the technical problems of this application, and achieve the expected technical effects of this application.

[0038] In this application, "preferred", "better", "more preferable", "preferably" are only used to describe the embodiments or examples with better effects, and it should be understood that they do not constitute a limitation on the protection scope of this application.

[0039] In this application, "further", "even further", "especially", etc. are used for descriptive purposes, indicating differences in content, but should not be understood as a limitation on the protection scope of this application.

[0040] In this application, "optionally", "optional", "option" mean that it can be either present or absent, that is, it refers to any one of the two parallel options of "present" or "absent". If "optional" appears multiple times in a technical solution, without special instructions and without contradictions or mutual restrictions, each "optional" is independent of each other.

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

[0042] In this application, for the technical features described in an open-ended manner, it includes a closed technical solution composed of the listed features, and also includes an open-ended technical solution containing the listed features.

[0043] In this application, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the selectable numerical values are considered continuous within the above-mentioned numerical intervals, and include the two numerical endpoints of the numerical range (i.e., the minimum value and the maximum value), as well as each numerical value between these two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to the integers within the numerical interval, it includes the two endpoint integers of the numerical range, as well as each integer between the two endpoints. In this article, it is equivalent to directly listing each integer. For example, when t is an integer selected from 1 to 10, it means that t is any integer selected from the integer group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In addition, when multiple range descriptions of features or characteristics are provided, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed in this article should be understood to include any and all sub-ranges subsumed therein.

[0044] For the temperature parameter in this application, unless otherwise specified, it allows both constant temperature treatment and variation within a certain temperature range. It should be understood that the constant temperature treatment allows the temperature to fluctuate within the accuracy range controlled by the instrument. It is allowed to fluctuate within a range such as ±5°C, ±4°C, ±3°C, ±2°C, ±1°C.

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

[0046] All documents mentioned in this application are cited as references in this application, just as if each document was cited separately as a reference. Unless it conflicts with the application purpose and / or technical solution of this application, the cited documents involved in this application are cited for all contents and all purposes. When this application involves cited documents, the definitions of relevant technical features, terms, nouns, phrases, etc. in the cited documents are also cited. When this application involves cited documents, the examples and preferred methods of the relevant technical features cited can also be included as references in this application, but only to the extent that this application can be implemented. It should be understood that when the cited content conflicts with the description in this application, this application shall prevail or be amended adaptively according to the description in this application.

[0047] In the first aspect of the embodiments of this application, a composition is provided. The composition includes glycine and trehalose with 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. Using the protein control product matrix solution added with this composition to prepare the CHI3L1 control product, the performance is stable. In particular, the control product does not need to be freeze-dried (i.e., stored in liquid state), and remains stable even after being opened and stored again, which can meet the current detection 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 includes glycine and trehalose with a weight ratio of (19.5 - 20.5):(9.5 - 10.5).

[0051] In the second aspect of the embodiments of this application, a protein control product matrix solution is provided, and the protein matrix solution includes the composition described above.

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

[0053] First, for the CHI3L1 quality control product of this application, by using animal serum, under the condition of high accuracy, the cost is greatly reduced, and it can be prepared in batches to meet the application of clinical clients.

[0054] Second, the CHI3L1 quality control product of this application can be in liquid form, which is beneficial for subsequent detection and convenient for various hospitals and testing institutions to verify and evaluate. At the same time, the expiration date after opening of the CHI3L1 quality control product of this application is much longer than that of the current freeze-dried products on the market, and its performance can still excellently meet the market demand.

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

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

[0057] In some examples of the present application, the protein control product matrix solution further includes a preservative. The present application does not particularly limit the type of the preservative, including but not limited to: PC300. The present application does not particularly limit the dosage of the preservative. An appropriate amount of the preservative can be added to the protein control product matrix solution, including but not limited to: the weight of the preservative corresponding to each 1 L of the fetal bovine serum is 0.3 g to 0.5 g (for example, 0.3, 0.35, 0.4, 0.45, 0.5 g).

[0058] In a third aspect of the embodiments of the present application, there is provided a CHI3L1 control product, which includes:

[0059] the composition or the protein control product matrix solution described above; and,

[0060] CHI3L1.

[0061] In some examples of the present application, the CHI3L1 control product is a combined product, and the combined product includes a plurality of control products and the concentrations of CHI3L1 in the plurality of control products are different. The present application does not particularly limit the control products, for example, 2, 3, 4, 5, 6, 7.

[0062] In some examples of the present application, the combined product includes two control products. Among them, the control product with a relatively high concentration of CHI3L1 can be called a high-concentration control product, and the control product with a relatively low concentration can be called a low-concentration control product.

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

[0064] a first-concentration control product, in which the concentration of CHI3L1 is 30 ng / mL to 50 ng / mL (for example, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50 ng / mL); and,

[0065] a second-concentration control product, in which the concentration of CHI3L1 is 150 ng / mL to 250 ng / mL (for example, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250 ng / mL).

[0066] In a fourth aspect of the embodiments of the present application, there is provided a CHI3L1 detection kit, and the CHI3L1 detection kit includes the CHI3L1 control product described above.

[0067] In a fifth aspect of the embodiments of the present application, there is provided a method for detecting CHI3L1 in a sample. The detection method uses the CHI3L1 control product or the CHI3L1 detection kit described above during the process of detecting CHI3L1 in the sample.

[0068] The implementation solutions of the present application will be described in detail below in conjunction with the embodiments. It should be understood that these embodiments are only used to illustrate the present application and not to limit the scope of the present application. For the experimental methods without specific conditions noted in the following embodiments, the guidelines given in the present application are preferentially referred to, and it is also possible to follow the experimental manuals or conventional conditions in the art, or the conditions recommended by the manufacturer, or refer to the experimental methods known in the art.

[0069] In the following specific embodiments, regarding the measurement parameters of the raw material components, if there is no special indication, there may be slight deviations within the weighing accuracy range. Regarding the temperature and time parameters, acceptable deviations caused by instrument test accuracy or operation accuracy are allowed.

[0070] The following embodiments are further descriptions of the present invention rather than limitations on the present invention. Unless otherwise specified, the equipment and reagents used in the present invention are conventional commercially available products in this technical field.

[0071] Example 1: Adding trehalose and glycine to the quality control product matrix solution effectively improves the stability of the quality control product for this item.

[0072] The quality control product matrix solution is divided into two parts, numbered 1# and 2# respectively. Among them, no excipients are added to the matrix solution of 1#, and the matrix solution consists of fetal bovine serum and PC300. The dosage of PC300 corresponding to every 1 L of fetal bovine serum is 0.5 g, while a certain proportion of trehalose and glycine are added to the matrix solution of 2#. The matrix solution consists of fetal bovine serum, glycine, trehalose and PC300. The dosages of glycine, trehalose and PC300 corresponding to every 1 L of fetal bovine serum are 20 g, 10 g and 0.5 g, denoted as 2#'.

[0073] The 1# is divided into two parts of the quality control product matrix solution, numbered 3# and 4# respectively; the 2#' is divided into two parts of the quality control product matrix solution, numbered 5# and 6# respectively.

[0074] An equal and low amount of CHI3L1 antigen is added to the above 3# and 5# to form CHI3L1 quality control products 3# and 5#.

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

[0076] They are respectively sub-packed in medium-borosilicate transparent glass bottles according to the specification of 1 mL / bottle. Three replicates are set for each detection condition, and the average value is taken correspondingly for subsequent tests.

[0077] Detect the CHI3L1 antigen concentration after 10 days of thermal acceleration at 37°C, 10 days at 2°C - 8°C after freeze-drying and reconstitution, 10 days at -20°C, 10 days at -80°C, and 3 days of storage at 25°C respectively, and calculate the relative change rate.

[0078] The concentration of CHI3L1 antigen in the CHI3L1 quality control product was measured using a chitinase 3-like protein 1 assay kit (chemiluminescence method) with a YahuiLong chemiluminescence analyzer. The test results and the corresponding calculated relative change rates are shown in Table 1.

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

[0080] Quality control product solution Initial concentration of CHI3L1 antigen 37°C - 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] From Table 1, for the CHI3L1 quality control product, comparing the results of No. 3 and No. 5, and No. 4 and No. 6 respectively. The quality control test results of No. 5 and No. 6 show that adding glycine and trehalose to the matrix solution can effectively improve the thermal stability of the quality control product compared to directly using the matrix solution of the quality control product. Among them: after the quality control product is treated at 37°C for 10 days, the relative deviation of No. 3 quality control product is -11.25%, and the relative deviation of No. 5 quality control product is 1.83%; the relative deviation of No. 4 quality control product is -14.97%, and the relative deviation of No. 6 quality control product is -5.48%. It can be seen that adding a certain proportion of trehalose and glycine to the matrix solution of the quality control product can effectively improve the thermal stability of the quality control product.

[0082] Example 2: Preparation process of chitinase 3-like protein 1 quality control product

[0083] A preparation method of a chitinase 3-like protein 1 quality control product includes the following steps:

[0084] (1) Raw material treatment and concentration confirmation: Measure the concentration of the CHI3L1 antigen raw material using a magnetic particle chemiluminescence detection kit.

[0085] (2) Preparation of the quality control product matrix solution: Prepare the quality control product matrix solution in a production workshop of 100,000-class cleanliness. The preparation process is as follows: Taking 1L of the quality control product matrix solution as an example, calculate the weighing amount according to the content of each component (2wt% glycine + 1wt% trehalose + 0.05wt% PC300). Then prepare 1L of fetal bovine serum solution, and sequentially add the weighed PC-300, glycine, and trehalose to the fetal bovine serum, and stir and mix for 15 - 20 minutes until the solution is clear and transparent.

[0086] (3) Preparation of the quality control product: Add the appropriate amount of the above antigen to the quality control product matrix solution, detect the level of each antigen using the magnetic particle chemiluminescence method, and perform labeling to obtain the following concentration quality control product combinations;

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

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

[0089] The above-mentioned quality control products at level 1 and level 2 are respectively tested. If the test results meet the following quality standards, the next step of sub-packaging will be carried out; if the test results do not meet the following standards, the concentration value can be adjusted by adding chitinase 3-like protein 1 antigen until the following quality standards are met;

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

[0091] (4) Sub-packaging of quality control products

[0092] The above-mentioned quality control products are sub-packaged into selected clean glass bottles at a specification of 2 mL per vial using an electric continuous dispenser, and rubber stoppers are added.

[0093] Example 3: Verification of the stability of chitinase 3-like protein 1 quality control products

[0094] 1. Thermal accelerated stability

[0095] To verify the performance of the above-mentioned chitinase 3-like protein 1 quality control products in extreme environments such as high temperature, the quality control products that are opened and then closed in this example are heat-treated at 37 °C for 11 days, and the unopened quality control products (shelf life of 18 months) stored at 2 °C - 8 °C are used as a control.

[0096] A chitinase 3-like protein 1 (CHI3L1) detection kit (chemiluminescence method) is selected to measure the luminescence value and concentration value of the markers of the above-mentioned chitinase 3-like protein 1 quality control products under each condition on the chemiluminescence platform. The stability is good, and the relative deviation is within ±10%. The results are shown in the following table.

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

[0098]

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

[0100]

[0101] 2. Verification of the stability of chitinase 3-like protein 1 quality control product after opening

[0102] To determine the stability of the above chitinase 3-like protein 1 quality control product after opening (stored with the cap closed under sterile conditions) under different storage conditions, in this example, the quality control product was placed 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 after opening. The quality control product stored at 2 °C - 8 °C (unopened, with a shelf life of 18 months) was used as a control.

[0103] A chitinase 3-like protein 1 (CHI3L1) detection kit (chemiluminescence method) was selected to measure the luminescence value and concentration value of the marker of the above chitinase 3-like protein 1 quality control product under each condition on the chemiluminescence platform. The stability was good, and the relative deviation was within ±10%. The results are shown in the following table:

[0104] Table 4. Results of the stability of chitinase 3-like protein 1 quality control product (level 1) after opening

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

[0106] Table 5. Results of the stability of chitinase 3-like protein 1 quality control product (level 2) after opening

[0107]

[0108]

[0109] Example 4: Application of chitinase 3-like protein 1 quality control product

[0110] (1) Preparation of the quality control product: Take out the quality control product from the packaging box and equilibrate it to room temperature. Carefully open the bottle cap to avoid liquid splashing.

[0111] (2) Accuracy detection of the quality control product: On the chemiluminescence analyzer, take the above-mentioned shaken and mixed quality control product as a sample, detect the CHI3L1 item 3 times, calculate the average value, and calculate the deviation between the average value and the given target value to investigate the accuracy of the quality control product. The accuracy results all meet the requirement that the relative deviation is within ±10%. The results are shown in the following table.

[0112] Table 6. Data of the accuracy detection of chitinase 3-like protein 1 quality control product

[0113]

[0114] Table 6 results showed that the deviations of each item of chitinase 3 protein 1 control product at level 1 and level 2 were -0.54% and -3.37% respectively, indicating that the deviation between the measured value of the control product and the calibrated target value did not exceed 10%, and the accuracy of the control products at level 1 and level 2 met the standards.

[0115] Example 5:

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

[0117] Table 7

[0118]

[0119] Secondly, referring to Example 3, perform the stability test, and the results are as follows:

[0120] Table 8, Accelerated Stability Results of Chitinase 3-Like Protein 1 Combined 1-4 Control Products (Level 1)

[0121]

[0122] Table 9, Accelerated Stability Results of Chitinase 3-Like Protein 1 Combined 1-4 Control Products (Level 2)

[0123]

[0124] Table 10, Open Stability Results of Chitinase 3-Like Protein 1 Combined 1-4 Control Products (Level 1)

[0125]

[0126]

[0127]

[0128] Table 11, Open Stability Results of Chitinase 3-Like Protein 1 Combined 1-4 Control Products (Level 2)

[0129]

[0130]

[0131] Comparative Example 1:

[0132] First, referring to Example 2, prepare the CHI3L1 control product samples, where

[0133] Control control product combination 1, including control control product 1-low and control control product 1-high, is different from the control products at level 1 and level 2 in Example 2 in that arginine is used instead of glycine.

[0134] Control quality control product combination 2, including control quality control product 2 - low and control quality control product 2 - high, is different from the quality control product at level 1 and the quality control product at level 2 in Example 2 in that sucrose is used instead of trehalose.

[0135] Control quality control product combination 3, including control quality control product 3 - low and control quality control product 3 - high, is different from the quality control product at level 1 and the quality control product at level 2 in Example 2 in that the concentration of glycine corresponding to every 1 L of fetal bovine serum is 21.5 g.

[0136] Control quality control product combination 4, including control quality control product 4 - low and control quality control product 4 - high, is different from the quality control product at level 1 and the quality control product at level 2 in Example 2 in that the concentration of trehalose corresponding to every 1 L of fetal bovine serum is 12 g.

[0137] Secondly, refer to Example 3 for the stability test, and the results are as follows:

[0138] Table 12. Accelerated stability results of chitinase 3 - like protein 1 control quality control combinations 1 - 4 (level 1)

[0139]

[0140] Table 13. Accelerated stability results of chitinase 3 - like protein 1 control quality control combinations 1 - 4 (level 1)

[0141]

[0142]

[0143] Table 14. Open - vial stability results of chitinase 3 - like protein 1 control quality control combinations 1 - 4 (level 1)

[0144]

[0145]

[0146]

[0147] Table 15. Open - vial stability results of chitinase 3 - like protein 1 control quality control combinations 1 - 4 (level 2)

[0148]

[0149]

[0150] In summary, the quality control product of the present application involves important markers for the diagnosis of liver fibrosis, providing a more stable, convenient, and lower-cost quality control substance for liver fibrosis detection reagents. The chitinase 3-like protein 1 quality control product prepared in the present application uses the liquid refrigeration method to prepare the chitinase 3-like protein 1 quality control into a liquid quality control, which can greatly facilitate the instrument operation and report output of small and medium-sized hospitals or testing institutions; the matrix liquid formula of the quality control product prepared in the present application can greatly improve the thermal stability and opening stability of CHI3L1, and even in terms of the shelf life, it is longer than that of the freeze-dried quality control on the market; the quality control product prepared by the present invention has a lower cost, better meets the commercial batch demand of the market, and ensures clinical application and laboratory testing efficiency, etc.

[0151] The technical features of the above-described embodiments and examples can be combined in any suitable manner. For the sake of brevity of description, 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, it should be considered to be within the scope described in this specification.

[0152] The above-described embodiments only represent several implementation manners of the present application, which are convenient for understanding the technical solutions of the present application specifically and in detail, but should not be construed as a limitation on the scope of patent protection of the application. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present application, several modifications and improvements can be made, and these all belong to the protection scope of the present application. In addition, it should be understood that after reading the above teachings of the present application, those skilled in the art can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the protection scope of the present application. It should also be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning, or limited experiments based on the technical solutions provided in the present application are all within the protection scope of the appended claims of the present application. Therefore, the protection scope of the patent of the present application shall be subject to the content of the appended claims, and the specification can be used to explain the content of the claims.

Claims

1. A composition, characterized in that The composition comprises glycine and trehalose in a weight ratio of (19-21):(8.5-11.5).

2. The composition according to claim 1, characterized in that The composition comprises glycine and trehalose in a weight ratio of (19.5-20.5):(9.5-10.5).

3. A protein control matrix solution, characterized in that: The protein matrix liquid comprises the composition according to any one of claims 1 to 2.

4. The protein control matrix solution according to claim 3, characterized in that The protein control matrix solution includes glycine, trehalose and fetal bovine serum; Optionally, the weights of glycine and trehalose corresponding to each 1L of the fetal bovine serum are 19 g to 21 g and 8.5 g to 11.5 g, respectively; Optionally, the weights of glycine and trehalose corresponding to each 1L of the fetal bovine serum are 19.5 g to 20.5 g and 9.5 g to 10.5 g, respectively.

5. The protein control matrix solution according to claim 4, characterized in that: The protein control matrix solution also includes a preservative; Optionally, the preservative includes PC300; Optionally, the weight of the preservative corresponding to each 1L of fetal bovine serum is 0.3g to 0.5g.

6. A CHI3L1 quality control product, characterized in that: The CHI3L1 quality control products include: The composition according to any one of claims 1 to 2 or the protein control matrix solution according to any one of claims 3 to 5; and CHI3L1.

7. The CHI3L1 quality control product according to claim 6, characterized in that: The CHI3L1 quality control product is a combination product, which includes multiple quality control products and the concentrations of CHI3L1 in the multiple quality control products are different.

8. The CHI3L1 quality control product according to claim 7, characterized in that: The combination product includes two quality control products; optionally, the combination product includes: A first concentration quality control product, wherein the concentration of CHI3L1 is 30 ng / mL to 50 ng / mL; and The second concentration quality control product, wherein the concentration of CHI3L1 is 150ng / mL to 250ng / mL.

9. A CHI3L1 detection kit, characterized in that: The CHI3L1 detection kit comprises the CHI3L1 quality control product according to any one of claims 6 to 8.

10. A method for detecting CHI3L1 in a sample, characterized in that: The detection method uses the CHI3L1 quality control product described in any one of claims 6 to 8 or the CHI3L1 detection kit described in claim 9 during the detection of CHI3L1 in the sample.

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