Hydroformylated red blood cell and whole blood quality control product

Abstract

The invention relates to the field of biological medicine, in particular to a whole blood quality control product which comprises mixed hydroformylated red blood cells, protection liquid and plasma. The preparation method of the hydroformylated red blood cells comprises the following steps: pre-immobilizing red blood cells through non-crosslinked aldehyde, and then mainly immobilizing the red blood cells through crosslinked aldehyde; a quality control standard substance is also added into the whole blood quality control product, and the quality control standard substance is one or more of cholesterol, triglyceride, high-density lipoprotein, low-density lipoprotein, blood sugar and uric acid. Compared with the prior art, the whole blood quality control product has the advantages of low red blood cell hemolysis rate and longer shelf life.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, in particular to an aldehyde-treated red blood cell and whole blood quality control product. BACKGROUND

[0002] The whole blood quality control product is a standardized product simulating the composition of human whole blood, which mainly includes red blood cells, plasma, quality control standards, protective solution and other components. It is widely used in blood detection projects such as blood routine detection, blood biochemical analysis, and coagulation function determination in clinical examination. By regularly detecting the quality control product, it helps the laboratory to verify the performance of the detection equipment, calibrate the detection method, and evaluate the technical level of the operator, so as to ensure the accuracy and consistency of the clinical detection results.

[0003] At present, in order to prolong the shelf life, the red blood cells in the whole blood quality control product are usually aldehyde-fixed red blood cells, commonly known as aldehyde-treated red blood cells. Aldehyde and the proteins on the surface and inside of the red blood cells undergo covalent cross-linking reaction, which makes the structure of the red blood cells fixed, the biological activity of the proteins on the surface and inside of the cells lost, but the morphology and antigenicity retained.

[0004] However, the shelf life of the current whole blood quality control product, especially the aldehyde-treated red blood cells, is still relatively short. After being preserved for a period of time, the phenomenon of red blood cell hemolysis, i.e. cell rupture and release of cell contents into the plasma, easily occurs. The released red blood cell contents can react with other substances in the whole blood quality control product, resulting in inaccurate detection results of biochemical indicators. SUMMARY

[0005] Therefore, the purpose of the present application is to overcome the defects or deficiencies of the prior art, and to provide an aldehyde-treated red blood cell.

[0006] An aldehyde-treated red blood cell, which is obtained by pre-fixing red blood cells with a non-cross-linking aldehyde and then main-fixing with a cross-linking aldehyde.

[0007] Compared with the prior art, the aldehyde-treated red blood cell provided by the present application avoids excessive aldehyde fixation, reduces the hemolysis rate, and can prolong the shelf life.

[0008] In an embodiment, the non-cross-linking aldehyde is an aldehyde containing only one aldehyde group in the molecule.

[0009] In an embodiment, the molecule of the non-cross-linking aldehyde further includes at least one hydroxyl group.

[0010] In an embodiment, the non-cross-linking aldehyde is DL-glyceraldehyde.

[0011] In an embodiment, the cross-linking aldehyde is glutaraldehyde.

[0012] In an embodiment, the method of pre-fixing is: resuspending the red blood cells in a non-crosslinking aldehyde solution with a concentration of 1 mM-20 mM, and incubating to pre-fix.

[0013] In an embodiment, the method of main fixing is: adding a crosslinking aldehyde solution with a final concentration of 0.01-1% v / v to the mixture of non-crosslinking aldehyde and red blood cells, and incubating to main fix.

[0014] In an embodiment, after the main fixing is completed, the non-crosslinking aldehyde and the crosslinking aldehyde solution are removed to obtain aldehyde-treated red blood cells, and the aldehyde-treated red blood cells are resuspended in a preservation solution.

[0015] The present application also provides a whole blood quality control product, comprising mixed aldehyde-treated red blood cells, a protective solution, and plasma, wherein the aldehyde-treated red blood cells are as described above; and the whole blood quality control product further comprises a quality control standard, which is one or more of cholesterol, triglyceride, high-density liprotein, low-density liprotein, blood glucose, and uric acid. Compared with the prior art, the hemolysis rate of the red blood cells in the whole blood quality control product is low, and the shelf life is longer.

[0016] In an embodiment, the hematocrit of the aldehyde-treated red blood cells in the whole blood quality control product is 5-35%.

[0017] In an embodiment, the volume ratio of the protective solution to the plasma is 5-15: 50-90.

[0018] In an embodiment, the preservation solution is a solution of trehalose, a preservative, an antioxidant, and bovine serum albumin added to a PBS aqueous solution.

[0019] In an embodiment, the whole blood quality control product further comprises a composite antioxidant, which comprises glutathione and biocystin. Through the antioxidant effect of glutathione and biocystin, the oxygen partial pressure in the whole blood quality control product can be controlled, so that the whole blood quality control product is suitable for both dry chemical detection and electrochemical detection. In addition, the antioxidant effect can also delay the degradation of the quality control standard, further prolonging the shelf life. DETAILED DESCRIPTION

[0020] This invention analyzes the reasons why aldehyde-fixed erythrocytes in current whole blood quality control products are prone to hemolysis. This invention finds that the aldehydes currently used for aldehyde fixation of erythrocytes are all cross-linked aldehydes. Cross-linked aldehydes generally refer to compounds containing two or more active aldehyde groups, or compounds that can form multi-site cross-linked structures during reactions with proteins. Cross-linked aldehydes form cross-linked structures by undergoing condensation reactions with amino groups of erythrocyte membrane proteins and cytoplasmic proteins, inhibiting intracellular physiological activities and enhancing the mechanical stability and chemical tolerance of erythrocytes, thereby achieving aldehyde fixation of erythrocytes. For example, commonly used glutaraldehyde can cross-link with erythrocyte membrane proteins through its dialdehyde group to form a stable network structure; as a special case, formaldehyde, although it has only one aldehyde group, can polymerize into paraformaldehyde and cross-link with various components of erythrocytes through multiple aldehyde group sites, achieving cell morphology fixation. In addition, some cross-linked aldehydes, such as acetaldehyde-glutaraldehyde and glutaraldehyde, in addition to cross-linking with proteins, can also inhibit intracellular physiological activities by inhibiting glycolysis in blood samples, which is equivalent to fixing erythrocytes.

[0021] To ensure the effective cross-linking of intracellular proteins by cross-linking aldehydes, a relatively high concentration is typically required, allowing them to enter the cytoplasm under sufficient osmotic pressure and act on intracellular proteins. However, high concentrations of cross-linking aldehydes can also lead to excessive cross-linking of cell membrane proteins, causing aldehyde-induced erythrocyte membrane hardening and morphological shrinkage (manifested as serrated edges), thus significantly increasing the hemolysis rate and severely affecting the long-term stability of the sample. Residual cross-linking aldehydes can also alter the properties of various substances within whole blood controls (RBCs), affecting the accuracy of the detection system. Furthermore, during testing, if there is significant hemolysis of erythrocytes in the whole blood control, it will affect the absorption rate and flow rate of the whole blood control on the test strip of the instrument. Simultaneously, the cellular contents released from ruptured erythrocytes also affect the overall ionic strength of the whole blood control, thus influencing the reaction kinetics during the detection process, shortening or lengthening the reaction time, and ultimately impacting the accuracy of the detection equipment.

[0022] In summary, there is an irreconcilable technical contradiction in the use of cross-linked aldehydes: ensuring sufficient cross-linking of intracellular proteins for aldehyde fixation and avoiding excessive cross-linking of cell membrane proteins to reduce hemolysis.

[0023] Therefore, this invention provides a novel method for erythrocyte aldehyde fixation, which involves pre-fixation with a non-crosslinked aldehyde followed by primary fixation with a crosslinked aldehyde. The non-crosslinked aldehyde is an aldehyde with a single number of aldehyde groups or limited reactivity, capable of binding to erythrocytes at a single site. It tends to reversibly bind to the amino groups of proteins, rather than undergoing irreversible covalent crosslinking with membrane proteins or intracellular structural proteins as crosslinked aldehydes do. This reversible binding can inhibit the activity of glycolytic enzymes (such as phosphofructokinase) within the cell, stabilizing intracellular glucose concentration and inhibiting various intracellular physiological activities. Simultaneously, it avoids excessive crosslinking of membrane proteins, maintaining the flexibility and biconcave disc morphology of erythrocytes. After pre-fixation with a non-crosslinked aldehyde, aldehyde fixation with a crosslinked aldehyde is less likely to result in excessive crosslinking, and extremely low concentrations of crosslinked aldehyde are sufficient to meet the erythrocyte aldehyde fixation requirements. Furthermore, this low concentration of crosslinked aldehyde allows for only mild crosslinking of the cell surface, enhancing the mechanical strength of the cell membrane and reducing the risk of hemolysis.

[0024] Furthermore, among numerous non-crosslinked aldehydes, this invention selected DL-glyceraldehyde for subsequent research. Because DL-glyceraldehyde has only one aldehyde group and two hydroxyl groups, the hydroxyl groups give it a certain degree of hydrophilicity. DL-glyceraldehyde reacts with some amino acids to form Schiff bases, but it does not "crosslink" with cell surface proteins, further preventing excessive crosslinking of the cell membrane during subsequent master fixation. In addition, it can be rapidly transported across the erythrocyte membrane to quickly inhibit glycolysis and maintain stable blood glucose concentration.

[0025] The novel erythrocyte aldehyde fixation method described above was used, and the formulation of the preservation solution for the erythrocytes was adaptively adjusted to effectively extend the shelf life of the aldehyde-fixed erythrocytes, thereby also extending the shelf life of whole blood quality control products containing the aldehyde-fixed erythrocytes.

[0026] Furthermore, the present invention also notes that currently, whole blood quality control products are only applicable to dry chemical detection or electrochemical detection alone, and there is no whole blood quality control product that is applicable to both detection methods.

[0027] Electrochemical methods are commonly used to detect components such as blood glucose and uric acid. This method achieves quantification by generating a current signal through ion conduction. It is sensitive to hematocrit (HCT), oxygen partial pressure, and ionic strength in the quality control sample, requiring a low-ion interference environment. Dry chemistry methods, on the other hand, are used for the detection of four lipid parameters (cholesterol, triglycerides, high-density lipoprotein, and low-density lipoprotein) and hemoglobin. They rely on colorimetric reactions, requiring stable red blood cell morphology and sufficient enzyme activity, and necessitate high ionic strength to maintain reaction system stability. Electrochemical methods need to avoid ion interference and HCT fluctuations, while dry chemistry methods need to ensure cell structure and enzyme activity. This application constructs a phosphate buffer system that can dynamically maintain and adjust ionic strength, adds trehalose to maintain osmotic pressure, reduces osmotic fragility of red blood cells to decrease hemolysis, and experimentally optimizes the parameter range of hematocrit. Additionally, glutathione and biocytin hydrochloride are added as a compound antioxidant to maintain the oxygen partial pressure in the whole blood quality control sample and avoid fluctuations in oxygen partial pressure. This improvement not only meets the requirements of low ion interference in electrochemical methods, but also adapts to the colorimetric requirements of dry chemical methods through membrane structure protection, enabling simultaneous quality control of multiple indicators such as blood glucose, uric acid, blood lipids, and hemoglobin.

[0028] The present invention will now be described in detail.

[0029] S1 Acquisition of Red Blood Cells and Plasma

[0030] After collecting human venous blood, EDTA-K2 anticoagulation was added to a final concentration of 1.5-2.2 mg / mL. After centrifuging 2000g of venous blood for 10 min, the middle leukocyte layer was removed, and the upper plasma layer and the lower red blood cell layer were collected separately.

[0031] The obtained plasma and red blood cells were filtered through a 10 μm pore size filter membrane (such as PALL). The filter further reduces the amount of residual white blood cells (<0.01%).

[0032] Additionally, in some modified embodiments, the red blood cells may be commercially available animal-derived red blood cells. Aldehyded red blood cells, used as a control, are not subjected to further aldehyde treatment.

[0033] S2 erythrocyte aldehyde fixation.

[0034] S201 Red Blood Cell Prefixation. Red blood cells obtained in S1 are mixed and resuspended in a deionized aqueous solution containing 1mM-20mM DL-glyceraldehyde, so that the hematocrit of red blood cells is between 5-20%. The mixture is incubated at room temperature for 90-140 minutes, preferably 120 minutes, for prefixation, and gently shaken every 5-15 minutes.

[0035] S202 Red Blood Cell Primary Fixation. Add 0.1M PBS solution of glutaraldehyde to the mixture of S201 pre-fixed red blood cells and DL-glyceraldehyde until the final concentration of glutaraldehyde is 0.01-1% v / v (preferably 0.05%). Incubate at room temperature for 15-30 minutes, preferably 20 minutes, to perform primary fixation, gently shaking every 5 minutes.

[0036] In this embodiment, the DL-glyceraldehyde is specifically dissolved in deionized water to a concentration of 1mM-20mM. The glutaraldehyde is specifically diluted with 0.1M PBS (pH 7.4) to a concentration of 0.01-10% for use. The dilution concentration can be adjusted freely by technicians according to actual conditions.

[0037] S203 washes away aldehyde reagents. Centrifuge the mixture of red blood cells and aldehyde reagents from S202 at 2000g for 10 minutes, remove the supernatant, and resuspend the red blood cells in PBS. Repeat this washing process three times to remove any residual aldehyde reagents. Finally, centrifuge again to remove the PBS supernatant, obtaining a precipitate of aldehyde-treated red blood cells for subsequent steps.

[0038] In some embodiments, aldehyde-fixed erythrocytes, after completing the S2 aldehyde fixation step, can be resuspended in a preservation solution in step S3A for long-term storage as a erythrocyte suspension. In other embodiments, they can be directly processed in step S3B to prepare a whole blood quality control before storage or use. Alternatively, in a preferred embodiment, after first forming a erythrocyte suspension in S3A and storing it for a period of time, the aldehyde-fixed erythrocyte precipitate is obtained again by centrifugation and washing, and then prepared into a whole blood quality control through step S3B for continued storage or use.

[0039] S3A resuspends the aldehyde-modified erythrocytes of S203 in the preservation solution to form a erythrocyte suspension.

[0040] Aldehyded erythrocytes from S203 were resuspended in a preservation solution to form a erythrocyte suspension. The preservation solution was a PBS aqueous solution (pH 7.2±0.1) containing 0.5-3% w / v trehalose to control osmotic pressure, 0.8-1.2% v / v Proclin 300 as a preservative, 1.5 U / ml superoxide dismutase (SOD) as an antioxidant, and 0.5-1% w / v bovine serum albumin (BSA) to reduce cell aggregation. The PBS was a standard formulation, and its concentration (i.e., the concentration of all phosphate groups) was adjusted to 1-30 mM; in this embodiment, 1 mM was preferred.

[0041] When aldehyde-treated erythrocytes are stored alone in erythrocyte suspension for a long period, the hematocrit of erythrocytes in the suspension is controlled at 5-10%; when they are prepared as whole blood quality control for long-term storage, the hematocrit of erythrocytes in the whole blood quality control is controlled at 5-35%. Meanwhile, the storage conditions are all 2-8°C protected from light, preferably 4°C.

[0042] S3B uses S203 aldehyde-treated red blood cells to prepare whole blood quality control products.

[0043] Aldinated erythrocytes obtained from S203, preservation solution, and plasma are mixed in a specific ratio. The hematocrit of aldehyde-containing erythrocytes in the whole blood quality control is strictly controlled to be 5-35%, more preferably 15-35%. Furthermore, the volume ratio of preservation solution to plasma is 5-15:50-90. This can be understood as follows: if the volume fractions of aldehyde-containing erythrocytes, preservation solution, and plasma are a, b, and c, respectively, then a is between 5 and 35, b + c = 100 - a, and b:c is 5-15:50-90.

[0044] Then, a compound antioxidant and a quality control standard are added to the mixture of the three to obtain whole blood quality control products.

[0045] The composite antioxidants are glutathione at a final concentration of 0.1-0.5% w / v and bio-cytokinin at a final concentration of 0.1-0.5% in the whole blood quality control sample. The quality control standards are one or more of the following substances: 3.0-10.0 mmol / L cholesterol, 0.5-5.0 mmol / L triglycerides, 0.8-2.5 mmol / L high-density lipoprotein, 1.5-5.0 mmol / L low-density lipoprotein, 2.0-15 mmol / L blood glucose, and 220-600 μmol / L uric acid (all concentrations refer to the final concentration in the whole blood quality control sample). In this embodiment, all of the above quality control standards are added. If the whole blood quality control sample is used for quantitative detection of hemoglobin, the aldehyde-treated erythrocytes used are dissolved and their mean globulin level is measured, and then the standard hemoglobin content in the whole blood quality control sample is calculated proportionally.

[0046] In addition, since the compound antioxidants and quality control products are concentrated liquids or powders, their impact on the volume of whole blood quality control products is minimal. Therefore, the volume of the compound antioxidants and quality control standards can be ignored.

[0047] After preparing the whole blood quality control sample, store it at 4°C in the dark for a long period of time.

[0048] Examples 1-4

[0049] Based on the above preparation steps, the concentrations of DL-glyceraldehyde and glutaraldehyde during aldehyde fixation of erythrocytes, and the volume ratio of aldehyde-fixed erythrocytes, preservation solution, and plasma in the whole blood quality control were further adjusted. Experiments 1-4 were conducted to compare and test the erythrocyte hemolysis rate and compatibility with different detection methods. In Examples 1-3, aldehyde-fixed erythrocytes were prepared into erythrocyte suspensions for erythrocyte hemolysis rate experiments. In Example 4, aldehyde-fixed erythrocytes were prepared into whole blood quality control samples and stored for 6 months before erythrocyte hemolysis rate and compatibility with different detection methods experiments were performed. Specific experimental parameters are shown in Tables 1 and 2.

[0050] Comparative Examples 1 and 2

[0051] The main difference between Comparative Examples 1 and 2 and the Examples lies in the method of erythrocyte aldehyde fixation: erythrocytes are fixed using only glutaraldehyde. The specific steps are as follows:

[0052] Red blood cells were fixed by monoaldehyde: The red blood cells obtained in S1 were resuspended in glutaraldehyde PBS solution to make the hematocrit 5-20%, and fixed by incubation at room temperature for 30 minutes. The cells were gently shaken every 5 minutes for monoaldehyde fixation.

[0053] After fixation, the erythrocytes fixed by single aldehyde were centrifuged at 2000g for 10 minutes with a mixture of aldehyde reagent and aldehyde solution. The supernatant was removed and the erythrocytes were resuspended in PBS and washed. The washing was repeated three times to remove the residual aldehyde reagent. Finally, the erythrocytes were centrifuged to remove the precipitate and obtain aldehyde-fixed erythrocytes.

[0054] In Comparative Example 1, the concentration of glutaraldehyde was 0.05%, and after obtaining aldehyde-treated erythrocytes, they were resuspended in the same preservation solution as in the example to form a erythrocyte suspension with a hematocrit of 8%, and were stored for a long time at 4°C in the dark.

[0055] In Comparative Example 2, the concentration of glutaraldehyde was 2%, and after obtaining aldehyde-treated erythrocytes, they were resuspended in the same preservation solution as in the example to form a erythrocyte suspension with a hematocrit of 10%, which was then stored for a long time at 4°C in the dark.

[0056] Please see Tables 1 and 2 for other specific experimental parameters.

[0057] Red blood cell hemolysis rate detection

[0058] After storing the red blood cell suspensions or whole blood quality control samples prepared in Examples 1-4 and Comparative Examples 1-2 for 6 months, the hemolysis rate of the red blood cells was detected and statistically analyzed.

[0059] Take a sample of the red blood cell suspension or whole blood quality control sample preserved in each example and comparative example, add it to a glass slide, cover it with a coverslip, and observe the morphology of the red blood cells under a microscope.

[0060] Qualitative assessment of red blood cell condition: Red blood cells with shrunken cell membranes, serrated edges, or ruptured cell membranes are indicative of hemolysis. Intact red blood cells with clearly visible biconcave disc-like structures are considered healthy.

[0061] The total number of red blood cells in the field of view of a statistical microscope, and the number of red blood cells exhibiting hemolysis, are calculated as follows: Hemolysis rate = (Number of red blood cells exhibiting hemolysis ÷ Total number of red blood cells) × 100%.

[0062] Compatibility testing of different detection methods:

[0063] After storing the whole blood quality control samples prepared in each Example 4 for 6 months, the blood glucose, uric acid, cholesterol, triglycerides, high-density lipoprotein, and hemoglobin in the whole blood quality control samples were detected using a ZD401 dry biochemical analyzer according to the instrument's instruction manual. The final concentrations of the added quality control standards and the final concentration of hemoglobin calculated from the proportion of aldehyde erythrocytes were used as reference concentrations for comparison. Example 4 was technically replicated 5 times.

[0064] This dry biochemical analyzer is an instrument compatible with both dry chemical and electrochemical methods. Blood glucose and uric acid are detected using electrochemical methods, while cholesterol, triglycerides, high-density lipoprotein, and hemoglobin are detected using dry chemical methods.

[0065] Table 1 shows the parameters and test results for each example and comparative example of long-term preservation of red blood cell suspension.

[0066]

[0067] Table 2 shows the parameters and test results for each example and comparative example of long-term storage of whole blood as a quality control sample.

[0068]

[0069] The test results showed that the erythrocyte suspensions or whole blood quality control samples prepared by the aldehyde fixation method of non-crosslinked aldehyde pre-fixation + crosslinked aldehyde main fixation provided by the present invention in Examples 1-4 had a hemolysis rate of less than or equal to 4% after 6 months of storage, while Comparative Examples 1 and 2 showed a hemolysis rate of greater than 5%.

[0070] Analysis revealed that, although Comparative Example 1 used an extremely low concentration of glutaraldehyde, it only used glutaraldehyde for primary fixation without pre-fixation. Under the microscope, a large number of aldehyde-treated erythrocytes were broken up, resulting in severe hemolysis. This was because the concentration of glutaraldehyde was too low, and the osmotic pressure was insufficient to allow it to enter the cytoplasm. Glutaraldehyde only acted on the cell membrane, leading to insufficient protein cross-linking within the erythrocytes while excessive cross-linking occurred on the cell membrane surface, causing erythrocyte rupture. In contrast, Comparative Example 2 used a normal concentration of glutaraldehyde, as is common in existing technologies, and only used glutaraldehyde for direct aldehyde fixation. Under the microscope, a large number of erythrocytes showed wrinkled cell membranes and serrated edges, and most of these cells ruptured, exhibiting hemolysis. This means that while using a high concentration of glutaraldehyde satisfies the requirements for protein cross-linking within the cells, it alters the characteristics of erythrocytes, leading to a higher hemolysis rate. In contrast, the lower hemolysis rate achieved in Examples 1-4 effectively avoids hemoglobin leakage due to red blood cell rupture, reduces background interference, and maintains the integrity of cell morphology and antigen structure in the sample, greatly improving the stability and reliability of the quality control products. This substantially extends the shelf life of whole blood quality control products, significantly improving their stability and reliability during long-term storage, and providing a longer-lasting and more accurate quality control solution for clinical testing, scientific research, and other fields.

[0071] This invention effectively ensures the fixation effect inside red blood cells while avoiding excessive cross-linking of the red blood cell membrane by first using non-cross-linked aldehyde for pre-fixation and then using a very low concentration of cross-linked aldehyde for primary fixation. This effectively overcomes the technical contradiction between ensuring sufficient fixation of intracellular proteins and avoiding excessive cross-linking of cell membrane proteins that alters red blood cell properties and reduces hemolysis rate.

[0072] Meanwhile, the whole blood quality control sample from Example 4, tested on the Zhongke ZD401 dry biochemical analyzer using different detection methods, showed similar values ​​to the reference concentrations for blood glucose, uric acid, cholesterol, triglycerides, and hemoglobin. This further demonstrates that the whole blood quality control sample prepared using the method of this invention remains compatible with two different detection methods even after a certain period of storage, exhibiting method versatility and high accuracy. This improved versatility of the whole blood quality control sample significantly reduces the cost and management burden on laboratories that would otherwise need to stock multiple quality control samples due to differences in instrument types. Furthermore, it standardizes quality control criteria across different testing platforms, reducing discrepancies in test results caused by differences in instrument principles.

[0073] In summary, this invention optimizes the aldehyde fixation method for erythrocytes by first pre-fixing with non-crosslinked aldehydes and then primarily fixing with crosslinked aldehydes. This avoids the defects of excessive aldehyde fixation in existing technologies, maintains the morphology and characteristics of erythrocytes, reduces hemolysis rate, and thus extends the shelf life of erythrocytes and whole blood quality control products prepared using these erythrocytes. Furthermore, the formulation of the preservation solution adapted to the aldehyde-fixed erythrocytes obtained using this method, along with the addition of a compound antioxidant, allows the prepared whole blood quality control products to be used by both dry chemical and electrochemical detection methods. In addition, the use of the compound antioxidant can further reduce the degradation of triglycerides, uric acid, and cholesterol, and extend the shelf life of the whole blood quality control products.

[0074] The terminology used in the embodiments of this application is for the purpose of describing specific embodiments only and is not intended to limit the embodiments of this application. Unless otherwise specified, experimental methods, especially those without specific conditions, are generally performed under conventional conditions. Materials and reagents used in the embodiments, unless otherwise specified, are commercially available. The term "and / or" as used herein refers to and includes any or all possible combinations of one or more of the associated listed items. In the description of this application, those skilled in the art will understand the specific meaning of the above terms in this application according to the specific circumstances. Unless otherwise specified, experimental methods, especially those without specific conditions, are generally performed under conventional conditions. Materials and reagents used in the embodiments, unless otherwise specified, are commercially available.

[0075] The embodiments described above are merely examples of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.

Claims

1. An aldehyde-treated erythrocyte, characterized in that: The aldehyde-modified erythrocytes are obtained by pre-fixing erythrocytes with non-crosslinked aldehydes and then performing primary fixation with crosslinked aldehydes.

2. The aldehyde-treated erythrocytes according to claim 1, characterized in that: The non-crosslinked aldehyde is an aldehyde whose molecule contains only one aldehyde group.

3. The aldehyde-treated erythrocytes according to claim 2, characterized in that: The non-crosslinked aldehyde molecule also includes at least one hydroxyl group.

4. The aldehyde-treated erythrocytes according to claim 1, characterized in that: The non-crosslinked aldehyde is DL-glyceraldehyde; the crosslinked aldehyde is glutaraldehyde.

5. The aldehyde-treated erythrocytes according to any one of claims 1-4, characterized in that: The pre-fixation method is as follows: red blood cells are resuspended in a non-crosslinked aldehyde solution containing 1mM-20mM and incubated for pre-fixation; The primary fixation method is as follows: add a cross-linked aldehyde solution with a final concentration of 0.01-1% v / v to a mixture of non-cross-linked aldehyde and erythrocytes, and incubate for primary fixation.

6. The aldehyde-treated erythrocytes according to claim 5, characterized in that: After primary fixation, the non-crosslinked aldehyde and crosslinked aldehyde solutions were removed to obtain aldehyde-treated erythrocytes, which were then resuspended in a preservation solution.

7. A whole blood quality control product, characterized in that: It includes a mixture of aldehyde-treated red blood cells, a protective solution, and plasma, wherein the aldehyde-treated red blood cells are the aldehyde-treated red blood cells as described in any one of claims 1-6; The whole blood quality control product also contains quality control standards, which are one or more of cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein, blood glucose, and uric acid.

8. The whole blood quality control product according to claim 7, characterized in that: The hematocrit of the aldehyde-treated erythrocytes in the whole blood quality control was 5-35%.

9. The whole blood quality control product according to claim 8, characterized in that: The volume ratio of the protective solution to the plasma is 5-15:50-90.

10. The whole blood quality control product according to claim 7, characterized in that: The preservation solution is a solution of PBS aqueous solution with added trehalose, preservatives, antioxidants, and bovine serum albumin; the whole blood quality control product also contains a compound antioxidant, which includes glutathione and bio-cytokinin.

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