An n-terminal atrial natriuretic peptide test kit calibrator buffer, calibrator and kit

Abstract

The application belongs to the technical field of diagnostic reagents, and particularly relates to a N-terminal atrial natriuretic peptide detection kit calibrator buffer, a calibrator and a kit. The N-terminal atrial natriuretic peptide detection kit calibrator buffer provided by the application has good stability for storing N-terminal atrial natriuretic peptide antigen mother liquor, avoids the freeze-drying process, reduces the process flow and cost of manufacturers, improves user experience without reconstitution, and thus solves the technical problem of excessive detection error caused by reconstitution of the freeze-dried powder calibrator in the prior art.

Description

Technical Field

[0001] This application belongs to the field of diagnostic reagent technology, and in particular relates to a calibrator buffer, calibrator and reagent kit for an N-terminal atrial natriuretic peptide detection kit. Background Technology

[0002] Currently, the main clinical method for diagnosing and assessing heart failure is by detecting N-terminal atrial natriuretic peptide.

[0003] The in vitro quantitative detection of N-terminal atrial natriuretic peptide (NNP) mainly relies on NNP diagnostic kits. These kits are used in conjunction with NNP calibrators. For example, Roche Diagnostics provides calibrators for its NNP diagnostic kits. However, these calibrators are lyophilized powders. Before use, these lyophilized powder calibrators require manual reconstitution with a certain volume of purified water. Some manufacturers even require transferring the calibrators to other suitable containers after reconstitution. This manual process not only introduces errors but also leads to batch-to-batch variations, resulting in a poor user experience. Furthermore, the lyophilization process increases the manufacturer's costs. Summary of the Invention

[0004] In view of this, this application provides a calibrator buffer, calibrator, and kit for an N-terminal atrial natriuretic peptide detection kit, which solves the technical problem that the lyophilized powder calibrator in the prior art needs to be reconstituted, resulting in excessive detection error.

[0005] The first aspect of this application provides a calibrator buffer for an N-terminal atrial natriuretic peptide detection kit, comprising salt, surfactant, preservative, protein stabilizer, and buffer solution.

[0006] Preferably, the protein stabilizer includes Xibao Biotechnology protein stabilizer DCL0009A, bovine serum albumin, and glycerol.

[0007] Preferably, the salt is sodium chloride or potassium chloride;

[0008] Preferably, the surfactant is at least one of polyoxyethylene sorbitan fatty acid ester, polyoxyethylene-8-octylphenyl ether, and lauryl alcohol polyoxyethylene ether;

[0009] Preferably, the preservative is at least one selected from ProClin, sodium azide, penicillin, streptomycin, gentamicin sulfate, and 5-bromo-5-nitro-1,3-dioxanepropanediol;

[0010] Preferably, the protein stabilizer further includes at least one of human serum albumin, casein, and gelatin;

[0011] Preferably, the buffer solution is at least one of PBS, TRIS, MES, HEPES, or MOPS buffer solutions.

[0012] Preferably, the salt is sodium chloride;

[0013] The surfactant is polyoxyethylene-8-octylphenyl ether;

[0014] The preservative is ProClin;

[0015] The buffer solution is a TRIS buffer solution.

[0016] Preferably, the surfactant polyoxyethylene sorbitan fatty acid ester is from the TW20, TW40 or TW80 series, the surfactant polyoxyethylene-8-octylphenyl ether is from the TX45, TX100, TX305 or TX405 series, and the surfactant lauryl polyoxyethylene ether is from the Brij23, Brij30, Brij35, Brij58 or Brij97 series.

[0017] The preservative ProClin is either ProClin 300 or ProClin 950 series.

[0018] Preferably, the calibrator buffer of the N-terminal atrial natriuretic peptide detection kit comprises the following proportions;

[0019] Salt content: 0.1–4.0 wt%

[0020] Surfactant 0.01~3.00 w%.

[0021] Preservatives: 0.05%~1w%;

[0022] Protein stabilizer 0.01~20.00 w%.

[0023] Buffer solution balance;

[0024] The buffer solution has a concentration of 0.01M to 0.1M and a pH of 5.0 to 9.0.

[0025] Preferably, the calibrator buffer of the N-terminal atrial natriuretic peptide detection kit comprises the following proportions;

[0026] Sodium chloride 0.9% w%

[0027] Polyoxyethylene-8-octylphenyl ether 0.1 w%;

[0028] ProClin300 0.1w%

[0029] Xibao Biotechnology Protein Stabilizer DCL0009A 2w%;

[0030] Bovine serum albumin 2w%;

[0031] 10w% glycerol;

[0032] Remaining TRIS buffer solution;

[0033] The TRIS buffer solution had a concentration of 0.02 M and a pH of 7.4.

[0034] A second aspect of this application provides a calibrator for an N-terminal atrial natriuretic peptide (ATP) assay kit, comprising an N-terminal ATP and a calibrator buffer solution for the NTP assay kit.

[0035] Preferably, the N-terminal atrial natriuretic peptide (ATP) assay kit calibrator includes a first N-terminal Atrial natriuretic peptide (ATP) assay kit calibrator and a second N-terminal Atrial natriuretic peptide (ATP) assay kit calibrator.

[0036] The concentration of the first N-terminal atrial natriuretic peptide assay kit calibrator is 135~150 pg / mL;

[0037] The concentration of the second N-terminal atrial natriuretic peptide assay kit calibrator is 2600~2800 pg / mL.

[0038] A third aspect of this application provides an N-terminal atrial natriuretic peptide (ATP) detection kit, comprising an N-terminal ATP detection reagent, an N-terminal ATP detection quality control sample, and a calibrator for the aforementioned N-terminal ATP detection kit.

[0039] It should be noted that since the calibration buffer provided in this application does not require manual reconstitution of the lyophilized powder, it is beneficial to the accuracy of the test results after long-term storage of the N-terminal atrial natriuretic peptide test kit when used with the N-terminal atrial natriuretic peptide test reagent and the N-terminal atrial natriuretic peptide test quality control sample.

[0040] In summary, this application provides a calibrator buffer, calibrator, and kit for an N-terminal atrial natriuretic peptide (NTP) assay kit. The NTP calibrator buffer, comprising salt, surfactant, preservative, protein stabilizer, and buffer solution, maintains the signal retention rate of the NTP calibrator for over 12 months at low temperatures (2-8°C) and for over 7 days at 37°C. This demonstrates that the NTP calibrator buffer provided in this application can maintain the stability of the NTP antigen stock solution over a long period. Furthermore, as a novel preservation process for NTP calibrators, it avoids the drawbacks of conventional NTP calibrators, which require lyophilization to obtain lyophilized powder. The reconstitution of lyophilized powder can lead to uncontrollable factors and reduced accuracy in clinical testing results, thus solving the technical problem of excessive detection errors caused by the reconstitution of lyophilized powder calibrators in existing technologies. Detailed Implementation

[0041] This application provides a calibrator buffer, calibrator, and kit for an N-terminal atrial natriuretic peptide detection kit, which solves the technical problem in the prior art where the lyophilized powder calibrator needs to be reconstituted, resulting in excessive detection error.

[0042] The technical solutions of this application are described clearly and completely below. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments in this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0043] Example 1

[0044] Given that existing N-terminal atrial natriuretic peptide (NTP) diagnostic kits use lyophilized powder as the calibrator, requiring manual reconstitution with purified water, which can easily lead to detection errors, Example 1 of this application provides a novel preservation process for NTP calibrators. This process preserves the NTP calibrator using a calibrator buffer solution. The buffer solution for the NTP test kit comprises salts, surfactants, preservatives, protein stabilizers, and a buffer solution. This buffer solution provides good stability for preserving NTP, allowing the calibrator to remain stable for 12 months at 2–8°C. This avoids the need for reconstitution of lyophilized powder prepared using conventional preservation methods, thus improving the accuracy of clinical test results. This solves the technical problem of excessive detection errors caused by the need for reconstitution of lyophilized powder calibrators in existing technologies. Furthermore, reducing the lyophilization process lowers the manufacturer's process flow and costs, and eliminating the need for reconstitution also improves the user experience.

[0045] Regarding the specific types of salts, surfactants, preservatives, protein stabilizers, and buffer solutions used in the calibration buffer for the N-terminal atrial natriuretic peptide diagnostic kit, the present application preferably uses sodium chloride or potassium chloride as the salt; the surfactant is at least one of polyoxyethylene sorbitan fatty acid ester, polyoxyethylene-8-octylphenyl ether, and polyoxyethylene lauryl ether; the preservative is at least one of ProClin, sodium azide, penicillin, streptomycin, gentamicin sulfate, and 5-bromo-5-nitro-1,3-dioxane propanediol; the protein stabilizer is at least one of Seebio protein stabilizer DCL0009A, bovine serum albumin, human serum albumin, casein, gelatin, and glycerol; the buffer solution is at least one of PBS, TRIS, MES, HEPES, or MOPS buffer solution.

[0046] Regarding the specific formulation of the calibration buffer for the N-terminal atrial natriuretic peptide diagnostic kit, the present application provides 29 formulations, as shown in Table 1.

[0047]

[0048] Table 1

[0049] In Table 1, A1 to A5 are respectively selected from PBS, TRIS, MES, HEPES, and MOPS buffer solutions; B1 to B6 are respectively selected from Seebio protein stabilizer (product number: DCL0009A), bovine serum albumin, human serum albumin, casein, gelatin, and glycerol protein stabilizer; C1 to C2 are respectively selected from sodium chloride and potassium chloride; D1 to D12 are respectively selected from TW20, TW40, TW80, TX45, TX100,

[0050] TX305, TX405, Brij23, Brij30, Brij35, Brij58, and Brij97; E1 to E7 are respectively selected from PC300, PC950, sodium azide, penicillin, streptomycin, gentamicin sulfate, and 5-bromo-5-nitro-1,3-dioxane propanediol.

[0051] Regarding the ratio of each component in the N-terminal atrial natriuretic peptide in the calibration buffer for the N-terminal atrial natriuretic peptide diagnostic kit provided by the present application, the present application preferably uses 0.1 to 4.0 w% salt, 0.01 to 3.00 w% surfactant, 0.05% to 1 w% preservative, 0.01 to 20.00 w% protein stabilizer, and the balance is the buffer solution; among them, the concentration of the buffer solution is 0.01 M to 0.05 M, the pH is 5.0 to 9.0, and the total amount of salt, surfactant, preservative, protein stabilizer, and buffer solution is 100 w%.

[0052] For the optimal formulation of the calibrator buffer for the N-terminal atrial natriuretic peptide diagnostic kit provided in this application, the preferred formulation comprises 0.9 w% sodium chloride, 0.1 w% polyoxyethylene-8-octylphenyl ether, 0.1 w% ProClin, 2 w% Siba Biotech protein stabilizer DCL0009A, 1 w% bovine serum albumin, 10 w% glycerol, with the remainder being TRIS buffer solution; wherein the buffer solution concentration is 0.02M, pH is 7.4, and the total amount of sodium chloride salt, polyoxyethylene-8-octylphenyl ether surfactant, ProClin preservative, Siba Biotech protein stabilizer DCL0009A, bovine serum albumin, glycerol protein stabilizer, and TRIS buffer solution is 100 w.

[0053] Example 2

[0054] Example 2 of this application provides a method for preparing calibrators for the N-terminal atrial natriuretic peptide (NAP) detection kit using the calibrator buffer solution described in Example 1, to evaluate whether the buffer solution provided in this application would lead to calibrator errors. The preparation method involves uniformly mixing the salt, surfactant, preservative, protein stabilizer, and buffer solution formulation described in Example 1 with the NAP antigen stock solution. The NAP detection kit calibrators are low-concentration (Cal1) and high-concentration (Cal2) NAP detection kit calibrators. The prepared low-concentration (Cal1) and high-concentration (Cal2) NAP detection kit calibrators are detected using a fully automated chemiluminescence immunoassay analyzer (iNova3000). The detected NAP luminescence values ​​are shown below.

[0055] As shown in Table 2-5.

[0056]

[0057] Table 2

[0058]

[0059] Table 3

[0060]

[0061] Table 4

[0062]

[0063] Table 5

[0064] As shown in Tables 2-5, the 10 groups of low-concentration N-terminal atrial natriuretic peptide (NAP) test kit calibrators (Cal1) prepared by uniformly mixing the buffer solution provided in Example 1 of this application with the NAP antigen stock solution showed little variation. Furthermore, the variation among multiple tests of any one group of low-concentration NAP test kit calibrators (Cal1) was also small. This indicates that the buffer solution provided in this application can be used with low-concentration (135~150 pg / mL) NAP test kit calibrators (Cal1) with minimal error in preparation with the NAP antigen stock solution. Simultaneously, the high-concentration NAP test kit calibrator (Cal2) also showed similar results. This indicates that the buffer solution provided in this application can also be used with high-concentration (2600~2800 pg / mL) NAP test kit calibrators (Cal2) with minimal error in preparation with the NAP antigen stock solution, and can be used to preserve NAP, demonstrating its potential as a buffer solution for NAP natriuretic peptide calibrators.

[0065] Experimental Example 1

[0066] Example 1 of this application provides a buffer solution for preparing the N-terminal atrial natriuretic peptide (NAP) assay kit calibrator to optimize the effect of the buffer formulation provided in this application on preserving NAP. The preparation method is to mix 1 w% BSA and 0.05 w% PC300, as well as different buffer solutions, with the NAP antigen stock solution to prepare the NAP assay kit calibrator, including low concentration (Cal1, 135~150 pg / mL) and high concentration NAP assay kit calibrator. Cal2 (2600~2800 pg / mL) was used to prepare low-concentration (Cal1) and high-concentration N-terminal atrial natriuretic peptide (NAP) assay kit calibrators (Cal2). The calibrators were stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the NAP diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The luminescence value results of the NAP were shown in Table 6.

[0067]

[0068] Table 6

[0069] As shown in Table 6, among the buffer solutions provided in this application, the TRIS buffer exhibits the best preservation effect on the N-terminal atrial natriuretic peptide (ATP) stock solution, with a high signal retention rate compared to other buffer solutions. Furthermore, this application has optimized the optimal concentration and pH value of the TRIS buffer. The preparation method involves uniformly mixing 1 w% BSA and 0.05 w% PC300, along with different concentrations or pH values ​​of TRIS buffer solutions, with the N-terminal ATP stock solution to prepare the N-terminal ATP detection kit calibrator, including low concentrations (Cal1, 135~150 pg / mL) and high concentrations of N-terminal ATP. The calibrator for the sodium peptide assay kit (Cal2, 2600~2800 pg / mL) was prepared. The low-concentration (Cal1) and high-concentration calibrator for the N-terminal atrial natriuretic peptide assay kit (Cal2) were stored at 37℃. Solutions containing the target protein were taken from day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the N-terminal atrial natriuretic peptide diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The luminescence value results of the detected N-terminal atrial natriuretic peptide are shown in Tables 7-8.

[0070]

[0071] Table 7

[0072]

[0073] Table 8

[0074] As can be seen from Table 7-8, when the concentration of the TRIS buffer solution is 0.02M and the pH is 7.4, the preservation effect of the N-terminal atrial natriuretic antigen stock solution is the best, and the signal retention rate is higher than that of other buffer solutions with different concentrations and pH values.

[0075] Experiment Example 2

[0076] Example 2 of this application provides a buffer solution for preparing the N-terminal atrial natriuretic peptide (NAP) assay kit calibrator to optimize the effect of the buffer formulation provided in this application on preserving NAP. The preparation method is to mix 1 w% BSA, 0.05 w% PC300, and different protein stabilizers with the NAP antigen stock solution to prepare the NAP assay kit calibrator, including low concentration (Cal1, 135~150 pg / mL) and high concentration NAP assay kit calibrator. Cal2 (2600~2800 pg / mL) was used to prepare low-concentration (Cal1) and high-concentration N-terminal atrial natriuretic peptide (NAP) assay kit calibrators (Cal2). The calibrators were stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the NAP diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The luminescence value results of the NAP were shown in Table 9.

[0077]

[0078] Table 9

[0079] As shown in Table 9, among the buffer solutions provided in this application, the Siba Biotech protein stabilizer DCL0009A, bovine serum albumin, and glycerol exhibit the best preservation effect on the N-terminal atrial natriuretic peptide (ATP) stock solution, with high signal retention rates compared to other protein stabilizers. Furthermore, this application optimized the optimal concentrations of Siba Biotech protein stabilizer DCL0009A, bovine serum albumin, and glycerol. The preparation method involves uniformly mixing 1 w% BSA and 0.05 w% PC300, along with different concentrations of Siba Biotech protein stabilizer DCL0009A, bovine serum albumin, and glycerol, with the N-terminal ATP calibrator, including low concentrations (C...). Cal1 (135~150 pg / mL) and high-concentration N-terminal atrial natriuretic peptide (NTP) calibrators (Cal2 (2600~2800 pg / mL)) were prepared and stored at 37°C. Solutions containing the target protein were taken on day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the NTP diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The luminescence value results of the detected NTP are shown in Table 10-12.

[0080]

[0081] Table 10

[0082]

[0083] Table 11

[0084]

[0085] Table 12

[0086] As can be seen from Table 10-12, when the concentrations of Xibao Biotech protein stabilizer DCL0009A, bovine serum albumin, and glycerol are 2 w%, 2 w% (including 1 w% BSA basal solution), and 10 w%, respectively, the preservation effect on the N-terminal atrial natriuretic antigen stock solution is the best, and the signal retention rate is higher than that of other concentrations of Xibao Biotech protein stabilizer DCL0009A, bovine serum albumin, and glycerol buffer solutions.

[0087] Experimental Example 3

[0088] Example 3 of this application provides a buffer solution for preparing the N-terminal atrial natriuretic peptide (NAP) assay kit calibrator to optimize the effect of the buffer formulation provided in this application on preserving NAP. The preparation method involves mixing 1 w% BSA, 0.05 w% PC300, and different salts with the NAP antigen stock solution to prepare the NAP assay kit calibrator, including low concentration (Cal1, 135~150 pg / mL) and high concentration (Ca1) NAP assay kit calibrator. The low-concentration (Cal1) and high-concentration N-terminal atrial natriuretic peptide (NAP) calibrators (Cal2) of the prepared kits were stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the NAP diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The luminescence value results of the detected NAP are shown in Table 13.

[0089]

[0090] Table 13

[0091] As can be seen from Table 13, when the salt is selected from sodium chloride and the concentration is 0.9 w%, the preservation effect of the N-terminal atrial natriuretic antigen stock solution is the best, and the signal retention rate is higher than that of other salt buffer solutions.

[0092] Experiment Example 4

[0093] Example 4 of this application provides a buffer solution for preparing the N-terminal atrial natriuretic peptide (NAP) assay kit calibrator to optimize the effect of the buffer formulation provided in this application on preserving NAP. The preparation method involves mixing 1 w% BSA, 0.05 w% PC300, and different surfactants with the NAP antigen stock solution to prepare the NAP assay kit calibrator, including low concentration (Cal1, 135~150 pg / mL) and high concentration NAP assay kit calibrator. Cal2 (2600~2800 pg / mL) was used to prepare low-concentration (Cal1) and high-concentration N-terminal atrial natriuretic peptide (NAP) assay kit calibrators (Cal2). The calibrators were stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the NAP diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The luminescence value results of the NAP were shown in Table 14.

[0094]

[0095] Table 14

[0096] As shown in Table 14, among the buffer solutions provided in this application, surfactant TX100 exhibits the best preservation effect on the N-terminal atrial natriuretic peptide (ATP) stock solution, with a higher signal retention rate than other surfactants. Furthermore, this application has optimized the optimal concentration of surfactant TX100. The preparation method involves uniformly mixing 1 w% BSA, 0.05 w% PC300, and different concentrations of surfactant TX100 with the N-terminal ATP stock solution to prepare the N-terminal ATP detection kit calibrator, including low concentrations (Cal1, 135~150 pg / mL) and high concentrations of N-terminal ATP. The calibrator of the N-terminal atrial natriuretic peptide (NNP) assay kit (Cal2, 2600~2800 pg / mL) was prepared. The low-concentration (Cal1) and high-concentration N-terminal atrial natriuretic peptide (NNP) assay kit calibrator (Cal2) were stored at 37℃. Solutions containing the target protein were taken on day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the N-terminal atrial natriuretic peptide diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The luminescence value results of the detected N-terminal atrial natriuretic peptide are shown in Table 15.

[0097]

[0098] Table 15

[0099] As can be seen from Table 15, when the concentration of surfactant TX100 is 0.1 w%, it has the best preservation effect on the N-terminal atrial natriuretic antigen stock solution, and the signal retention rate is higher than that of surfactant TX100 at other concentrations.

[0100] Experimental Example 5

[0101] Example 5 of this application provides a buffer solution for preparing calibrators for the N-terminal atrial natriuretic peptide (NAP) assay kit to optimize the preservation effect of the buffer formulation provided in this application. The preparation method involves uniformly mixing 1 w% BSA, 0.05 w% PC300, and different preservatives with the NAP antigen stock solution to prepare the NAP assay kit calibrators, including low concentration (Cal1, 135~150 pg / mL) and high concentration (Cal1, 135~150 pg / mL). Cal2 (2600~2800 pg / mL) was used to prepare low-concentration (Cal1) and high-concentration N-terminal atrial natriuretic peptide (NAP) assay kit calibrators (Cal2). The calibrators were stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the NAP diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The luminescence value results of the NAP were shown in Table 16.

[0102]

[0103] Table 16

[0104] As shown in Table 16, among the buffer solutions provided in this application, the preservative ProClin300 exhibits the best preservation effect on the N-terminal atrial natriuretic peptide (NNP) antigen stock solution, with a higher signal retention rate than other preservatives. Furthermore, this application has optimized the optimal concentration of the preservative ProClin300. The preparation method involves uniformly mixing 1 w% BSA and 0.05 w% PC300, along with different concentrations of the preservative ProClin300, with the NNP antigen stock solution to prepare the NNP detection kit calibrator, including low concentrations (Cal1, 135~150 pg / mL) and high concentrations of NNP. The calibrator of the N-terminal atrial natriuretic peptide (NAP) assay kit (Cal2, 2600~2800 pg / mL) was prepared. The low-concentration (Cal1) and high-concentration NAP calibrator (Cal2) were stored at 37℃. Solutions containing the target protein were taken on day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviation was calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the NAP natriuretic peptide diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The luminescence value results of the detected NAP natriuretic peptide are shown in Table 17.

[0105]

[0106] Table 17

[0107] As can be seen from Table 17, when the concentration of preservative ProClin300 is 0.1 w% (including 0.05 w% ProClin300 base solution), it has the best preservation effect on the N-terminal atrial natriuretic antigen stock solution, and the signal retention rate is higher than that of other concentrations of preservative ProClin300 buffer solution.

[0108] Experimental Example 6

[0109] Example 6 of this application provides a performance test of the N-terminal atrial natriuretic peptide (NAP) assay kit calibrator buffer and the NAP antigen stock solution, prepared by uniformly mixing the NAP assay kit calibrator and the commercially available lyophilized powder calibrator from Roche Diagnostics, to compare their effects on preserving NAP (37°C). The test method includes first reconstituted the lyophilized powder calibrator from Roche Diagnostics with the NAP assay kit calibrator, and then adding 0.9 w% sodium chloride, 0.1 w% polyoxyethylene-8-octylphenyl ether, and 0.1 w%... ProClin, 2 w% Siba Biotech protein stabilizer DCL0009A, 1 w% bovine serum albumin, 10 w% glycerol, and the balance being TRIS buffer solution; wherein the TRIS buffer solution concentration is 0.02M, pH 7.4, is mixed evenly with the N-terminal atrial natriuretic peptide antigen stock solution to prepare the N-terminal atrial natriuretic peptide assay kit calibrator. The prepared N-terminal atrial natriuretic peptide assay kit calibrator includes low concentration (Cal1, 135~150 pg / mL) and high concentration N-terminal atrial natriuretic peptide. The peptide assay kit calibrator (Cal2, 2600~2800 pg / mL), after preparation, the low concentration (Cal1) and high concentration N-terminal atrial natriuretic peptide assay kit calibrator (Cal2) were stored at 37℃. Solutions containing the target protein were taken from day 0 and day 7, respectively, and detected using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the N-terminal atrial natriuretic peptide diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The results are shown in Table 17.

[0110]

[0111] Table 18

[0112] As shown in Table 18, the lyophilized powder calibrators provided by Roche Diagnostics exhibit poor reconstitution stability. After 7 days, the signal retention value of the low-concentration (Cal1, 135~150 pg / mL) N-terminal atrial natriuretic peptide (NAP) assay kit calibrator was only 67%, while the signal retention value of the high-concentration NAP assay kit calibrator (Cal2, 2600~2800 pg / mL) was 78%. In contrast, the NAP assay kit calibrator buffer solution (0.9 w% sodium chloride, 0.1 w% polyoxyethylene-8-octylphenyl ether, 0.1 w%...) provided in this application... The reagent consisted of ProClin, 2 w% of Siba Biotech protein stabilizer DCL0009A, 1 w% bovine serum albumin, 10 w% glycerol, and the remainder was TRIS buffer solution. The TRIS buffer solution (0.02 M, pH 7.4) showed a signal retention rate of up to 95% for the low-concentration (Cal1, 135~150 pg / mL) N-terminal atrial natriuretic peptide (NAP) calibrator after 7 days, while the high-concentration NAP calibrator (Cal2, 2600~2800 pg / mL) showed a signal retention rate of 96%. This demonstrates that the NAP calibrator buffer solution provided in this application can maintain the stability of the NAP antigen stock solution for a long period, while avoiding the need for lyophilization to prepare lyophilized powder, which increases the manufacturer's production process and costs, and reduces the user experience.

[0113] Experimental Example 7

[0114] Example 7 of this application provides a calibrator for the N-terminal atrial natriuretic peptide (NTP) assay kit prepared by uniformly mixing the calibrator buffer and the NTP antigen stock solution. The effects of these two methods on preserving NTP (2-8°C) were compared. The detection method involved mixing 0.9 w% sodium chloride, 0.1 w% polyoxyethylene-8-octylphenyl ether, 0.1 w% ProClin, 2 w% Siba Biotech protein stabilizer DCL0009A, 1 w% bovine serum albumin, 10 w% glycerol, with the remainder being TRIS buffer solution. The TRIS buffer solution had a concentration of 0.02 M and a pH of 7.4. This mixture was then thoroughly mixed with the NTP antigen stock solution to prepare the NTP calibrator. The prepared NTP calibrator included low concentrations (Cal1, 135-150 pg / mL) and high concentrations of NTP. The peptide assay kit calibrator (Cal2, 2600~2800 pg / mL), after preparation, the low concentration (Cal1) and high concentration N-terminal atrial natriuretic peptide assay kit calibrator (Cal2) were stored at 37℃. Solutions containing the target protein were taken on day 0 and day 7, respectively, and detected using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the N-terminal atrial natriuretic peptide diagnostic kit NT-proANP (magnetic microparticle chemiluminescence method). The results are shown in Table 19.

[0115]

[0116] Table 19

[0117] As shown in Table 19, the calibrator buffer solution (0.9 w% sodium chloride, 0.1 w% polyoxyethylene-8-octylphenyl ether, 0.1 ... The reagent consists of ProClin, 2w% Siba Biotech protein stabilizer DCL0009A, 1w% bovine serum albumin, 10w% glycerol, and the remainder is TRIS buffer solution. The TRIS buffer solution concentration is 0.02M, pH 7.4. After 13 months, the low-concentration (Cal1, 135~150 pg / mL) N-terminal atrial natriuretic peptide (NAP) calibrator retained a signal value as high as 93%, while the high-concentration NAP calibrator (Cal2, 2600~2800 pg / mL) retained a signal value of 89%. This demonstrates that the NAP calibrator buffer solution provided in this application can maintain the stability of the NAP antigen stock solution for a long period, while avoiding the need for lyophilization to prepare lyophilized powder, which increases the manufacturer's production process and costs, and reduces the user experience.

[0118] The above embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of this application.

Claims

1. A calibrator for an N-terminal atrial natriuretic peptide (ATP) assay kit, characterized in that, Includes N-terminal atrial natriuretic peptide and N-terminal atrial natriuretic peptide assay kit calibrator buffer; The calibrator buffer solution of the N-terminal atrial natriuretic peptide detection kit is composed of the following proportions; Sodium chloride 0.9% w% Polyoxyethylene-8-octylphenyl ether 0.1 wt% ProClin 0.1% Xibao Biotechnology Protein Stabilizer DCL0009A 2w% Bovine serum albumin 1w% 10% glycerin Remaining TRIS buffer solution; The TRIS buffer solution had a concentration of 0.02 M and a pH of 7.

4.

2. The calibrator for the N-terminal atrial natriuretic peptide detection kit according to claim 1, characterized in that, The N-terminal atrial natriuretic peptide (ATP) assay kit calibrators include a first N-terminal Atrial natriuretic peptide (ATP) assay kit calibrator and a second N-terminal Atrial natriuretic peptide (ATP) assay kit calibrator. The concentration of the first N-terminal atrial natriuretic peptide assay kit calibrator is 135~150 pg / mL; The concentration of the second N-terminal atrial natriuretic peptide assay kit calibrator is 2600~2800 pg / mL.

3. A kit for detecting N-terminal atrial natriuretic peptide, characterized in that, The invention includes an N-terminal atrial natriuretic peptide (ATP) assay reagent, an N-terminal Atrial natriuretic peptide (ATP) assay quality control material, and a calibrator for an N-terminal Atrial natriuretic peptide (ATP) assay kit as described in claim 1 or 2.

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