Antibody storage solution and use thereof

Abstract

The application provides an antibody storage solution, through optimization of components such as S-allyl-L-cysteine, poloxamer 188, ascorbic acid, ProClin300, N-acetyl-L-histidine and the adding proportion, a new antibody stabilizer system which is free of animal sources, can be long-term stable at room temperature, has antibody structure protection and function maintenance, is suitable for IVD and industrial production, and has important research value and urgent application demand.

Description

Technical Field

[0001] This invention relates to the field of biology, and in particular to an antibody preservation solution and its application. Background Technology

[0002] Antibodies, as a core class of biological reagents, have wide and crucial applications in in vitro diagnostics, biopharmaceuticals, and scientific research testing. However, antibodies are highly susceptible to various structural and functional damages during routine storage and transportation, such as protein denaturation, misfolding, molecular aggregation, adsorption to container walls leading to loss of active ingredients, decreased antigen-binding capacity, and significantly reduced biological activity.

[0003] Current mainstream antibody preservation strategies still have many limitations: they rely on ultra-low temperature cold chains, resulting in high storage and transportation costs and stringent conditions, making it difficult to meet the needs of rapid on-site testing and remote applications; animal-derived protein stabilizer systems suffer from problems such as large batch variations of raw materials, potential pathogen contamination, and residual animal-derived components, making it difficult to meet the strict quality control standards for in vitro diagnostic reagents and pharmaceutical proteins; while traditional protective agent systems such as glycerol and sucrose only have a certain effect on short-term antibody stability, have poor long-term stability at room temperature, and have limited protective effects on antibody higher-order structure and biological activity, failing to achieve comprehensive stability protection.

[0004] In summary, existing antibody preservation technologies cannot simultaneously meet the requirements of safe, stable, convenient, and compliant industrial applications. Therefore, the research and development of a novel antibody stabilizer system that is animal-free, stable at room temperature for extended periods, provides both antibody structural protection and functional maintenance, and is suitable for IVD and industrial production is of significant research value and urgently needed for application. Summary of the Invention

[0005] To address the problems existing in the prior art, the present invention provides an antibody preservation solution and its application.

[0006] The first aspect of the present invention provides an antibody preservation solution, comprising:

[0007] Non-animal-derived structure protectants with a weight-to-volume ratio (w / v) of 0.5%-10%;

[0008] An interfacial protectant with a mass-to-volume ratio (w / v) of 0.01%-0.2%;

[0009] Antioxidants with a w / v ratio of 0.01%-0.5%;

[0010] Preservatives with a w / v ratio of 0.01%-0.2%;

[0011] 10mM-50mM buffer solution;

[0012] The non-animal-derived structure protectant includes S-allyl-L-cysteine. Preferably, the interface protectant includes poloxamer 188.

[0013] Preferably, the antioxidant includes ascorbic acid.

[0014] Preferably, the preservative includes ProClin300.

[0015] Preferably, the buffer solution is a TPS buffer system. More preferably, the pH value of the buffer solution is 6.5-7.5.

[0016] Preferably, the antibody preservation solution further includes an anti-aggregation synergist at a weight-to-volume ratio (w / v) of 0.1%-2%. More preferably, the anti-aggregation synergist includes N-acetyl-L-histidine.

[0017] A second aspect of the present invention provides a method for preserving antibodies, comprising:

[0018] Add the antibody preservation solution as described in the first aspect of the present invention to the antibody sample to be preserved, and seal for preservation.

[0019] A third aspect of the present invention provides an antibody preservation kit, comprising the antibody preservation solution as provided in the first aspect of the present invention and the antibody preservation method as provided in the second aspect of the present invention.

[0020] The fourth aspect of the present invention provides the application of the antibody preservation solution as provided in the first aspect of the present invention, the antibody preservation method as described in the second aspect of the present invention, or the antibody preservation kit as described in the third aspect of the present invention in antibody preservation.

[0021] Compared with the prior art, the advantages of the present invention are:

[0022] 1. Animal-free characteristics: The antibody preservation solution provided by this invention does not contain any animal-derived components such as bovine serum albumin (BSA) or animal serum. This avoids the problems of batch variation and potential pathogen contamination caused by traditional animal-derived stabilizers from the source. It meets the strict quality control standards of in vitro diagnostics (IVD) and pharmaceutical fields, broadens the application scenarios of the preservation solution, and improves the safety and compliance of the product.

[0023] 2. Long-term stable at room temperature and significant practicality: This invention achieves long-term stable preservation of antibodies at room temperature (25±2℃) through a multi-dimensional synergistic protection system, with a preservation period of ≥6 months and an antibody activity retention rate of ≥90%. It eliminates the need for ultra-low temperature cold chain storage and transportation, significantly reducing storage and transportation costs. It solves the industry pain points of insufficient room temperature stability and reliance on cold chains in traditional preservation technologies, and is more suitable for industrial needs such as rapid on-site testing and remote application scenarios.

[0024] 3. Synergistic Protection Mechanism: This invention breaks through the traditional single protection mode and constructs a "dual synergistic protection" system with S-allyl-L-cysteine ​​and N-acetyl-L-histidine as the core, as detailed below:

[0025] (1) Long-term structural protection (S-allyl-L-cysteine): Based on the preferential exclusion effect, it can effectively stabilize the native conformation of the antibody, enhance the antibody's resistance to heat denaturation and hydrolysis, protect the antibody's active site, and provide a basic guarantee for the integrity of the antibody structure.

[0026] (2) Precise protection against aggregation (N-acetyl-L-histidine): Specifically shields the hydrophobic sites on the surface of antibody molecules and regulates the surface charge distribution, thus doubly inhibiting antibody molecule aggregation and reducing the loss of activity caused by aggregation from the root cause;

[0027] (3) Dual synergistic protection: While stabilizing the antibody conformation, SAC can enhance the binding ability of NAH to the hydrophobic sites on the surface of antibody molecules, thereby improving its anti-aggregation effect; while the aggregation-inhibiting effect of NAH can reduce the interference of abnormal aggregation of antibody molecules on the protective effect of SAC. The synergistic effect of the two can significantly improve the antibody activity retention rate after 6 months of storage at room temperature compared to using only one of the reagents, and the antibody activity retention rate under heat shock is even more significantly improved, effectively solving the pain points of traditional storage systems with single protection and poor synergy. Attached Figure Description

[0028] Figure 1 This is an image showing the immunocytochemical staining results of different antibody samples after 30 days of heat shock storage, as provided in an embodiment of the present invention; wherein... Figure 1 A is an immunocytochemical staining image of the antibody sample from Example 5 after heat shock. It can be seen that the target antigen sites in the cells show strong positive staining (brownish-yellow), the staining is uniform, the positive localization is accurate, the cell morphology is intact, the background is clean, and there is no obvious non-specific staining. The staining effect is close to that of fresh antibody, indicating that after storage at extreme high temperature, the antibody can still bind to the target antigen efficiently and the detection performance is stable. Figure 1 B is an immunocytochemical staining image of the fresh antibody positive control, which serves as the experimental benchmark. It shows that the target antigen sites in the cells show strong positive staining (brownish-yellow), with clear and uniform staining, accurate positive localization, no background interference, and intact cell morphology, which intuitively reflects the staining effect under normal antibody detection conditions. Figure 1 C is the immunocytochemical staining image of the antibody sample in Comparative Example 1. It can be seen that the cells have almost no positive staining, only weak non-specific light staining, and the background is messy, making it impossible to identify the target antigen site. Although the cell morphology is basically intact, the staining effect is extremely poor. Detailed Implementation

[0029] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0030] An antibody preservation solution comprising:

[0031] Non-animal-derived structure protectants with a weight-to-volume ratio (w / v) of 0.5%-10%;

[0032] An interfacial protectant with a mass-to-volume ratio (w / v) of 0.01%-0.2%;

[0033] Antioxidants with a w / v ratio of 0.01%-0.5%;

[0034] Preservatives with a w / v ratio of 0.01%-0.2%;

[0035] 10mM-50mM buffer solution;

[0036] The non-animal-derived structure protectant mentioned above includes S-allyl-L-cysteine.

[0037] In some embodiments, the interface protectant includes poloxamer 188.

[0038] In some embodiments, the antioxidant includes ascorbic acid.

[0039] In some embodiments, the preservative includes ProClin300.

[0040] In some embodiments, the buffer solution is a TPS buffer system. In some preferred embodiments, the pH value of the buffer solution is 6.5-7.5.

[0041] In some embodiments, the antibody preservation solution further includes an anti-aggregation synergist at a weight-to-volume ratio (w / v) of 0.1%-2%; in some preferred embodiments, the anti-aggregation synergist includes N-acetyl-L-histidine.

[0042] Example

[0043]

[0044] Application Example 1: Room Temperature Stability Test of Antibody Preservation Solution

[0045] 1. Experimental Objective

[0046] The preservation effect of the antibody preservation solution provided by this invention under long-term room temperature conditions was verified by room temperature stability test.

[0047] 2. Experimental Materials

[0048] 1) Antibody sample: IgG antibody (purity ≥95%), concentration 1 mg / mL;

[0049] 2) Test reagents: Antibody preservation solutions provided in Examples 1-3 of this invention, and existing conventional antibody preservation solutions (Comparative Example 1);

[0050] 3) Testing equipment: sterile centrifuge tubes, constant temperature incubator (accuracy ±0.5℃), ELISA reader, pipettes, etc.

[0051] 3. Experimental Methods

[0052] 3.1 Mix the IgG antibody with the antibody preservation solution provided in Examples 1-3 and Comparative Example 1 at a volume ratio of 1:10. After thorough mixing, dispense the mixture into sterile centrifuge tubes, seal them, and store them in a constant temperature incubator without any disturbance.

[0053] 3.2 Samples were taken at 2 months, 4 months and 6 months of storage, and equilibrated at room temperature for 30 minutes. Antibody activity was detected by enzyme-linked immunosorbent assay (ELISA). Three parallel replicates were set up for each time point and each sample, and the detection was performed three times. The average antibody activity of the three detection results was calculated. The antibody activity at the initial storage time was used as the benchmark to compare and calculate the activity retention rate at that time point.

[0054] 4. Experimental Results

[0055]

[0056] 5. Results Analysis

[0057] Experimental results showed that after 2, 4, and 6 months of storage at 25±2℃, the antibody activity retention rates of the groups using the antibody preservation solutions described in Examples 1-3 of this invention were significantly higher than those of Comparative Example 1. Example 2 showed the best results, with an antibody activity retention rate of 89.2±1.8% after 6 months of storage. Example 3 demonstrated stability during long-term storage, with an activity retention rate of 88.4±1.6% after 6 months. Even with the low concentration formulation of Example 1, the activity retention rate remained at 80.5±2.5% after 6 months, demonstrating significantly better overall stability than the existing conventional antibody preservation system of Comparative Example 1.

[0058] In contrast, in Comparative Example 1, which used the traditional glycerol + BSA system, the antibody activity showed a significant downward trend with prolonged storage time. The activity retention rate was only 81.9±1.6% after 2 months of storage, and further decreased to 64.2±2.4% after 6 months. Moreover, the detection deviation at each time point was large, indicating poor stability.

[0059] The above results indicate that the antibody preservation solution of the present invention, with S-allyl-L-cysteine ​​as the core protective component, can effectively slow down antibody denaturation and inactivation at room temperature, maintain the stability of antibody structure and function for a long time, and the overall preservation effect is significantly better than that of existing conventional antibody preservation solutions.

[0060] Application Example 2: Thermal Shock Resistance Test of Antibody Preservation Solution

[0061] 1. Experimental Objective

[0062] The thermal shock resistance of the antibody preservation solution provided by this invention was evaluated, and its long-term protective effect on antibodies was further demonstrated by simulating extreme room temperature or short-term high temperature storage scenarios.

[0063] To further enhance the long-lasting protective effect of the antibody preservation solution provided by this invention, N-acetyl-L-histidine is added based on Example 2. N-acetyl-L-histidine can specifically bind to hydrophobic sites on the surface of antibody molecules, shielding the hydrophobic interactions between antibody molecules. Simultaneously, it regulates the surface charge distribution of antibody molecules, reducing intermolecular electrostatic attraction and doubly inhibiting the tendency of antibody molecules to aggregate. This fundamentally reduces the loss of antibody activity caused by aggregation. Compared to traditional anti-aggregation reagents (such as arginine), its anti-aggregation effect is gentler and more durable, as detailed below:

[0064]

[0065] 2. Experimental Materials

[0066] 1) Antibody sample: IgG antibody (purity ≥95%), concentration 1 mg / mL;

[0067] 2) Test reagents: Antibody preservation solutions provided in Examples 1-3 of this invention, and existing conventional antibody preservation solutions (Comparative Example 1).

[0068] 3) Testing equipment: sterile centrifuge tubes, constant temperature incubator (accuracy ±0.5℃), ELISA reader, pipettes, etc.

[0069] 3. Experimental Methods

[0070] 3.1 Mix the IgG antibody with the antibody preservation solutions provided in Examples 2, 4-6, and Comparative Example 1 at a volume ratio of 1:10. After thorough mixing, dispense the mixture into sterile centrifuge tubes, seal them, and place them in a constant temperature incubator at 42°C for 7 days (simulating extreme room temperature and short-term high temperature storage scenarios, which are the usual and stringent conditions for thermal shock testing). Do not disturb the tubes during this period to ensure that the thermal shock conditions are stable.

[0071] 3.2 Samples were taken after 7 days of heat shock culture and placed at room temperature for 30 minutes to equilibrate. Antibody activity was detected by enzyme-linked immunosorbent assay (ELISA). Each sample was tested in 3 replicates. The average antibody activity of the 3 test results was calculated. The activity retention rate was calculated by comparing the antibody activity at the initial storage time.

[0072] 4. Experimental Results

[0073]

[0074] 5. Results Analysis

[0075] Experimental results showed that after 7 days of high-temperature shock at 42℃, the antibody activity retention rates of all the example groups of this invention were significantly higher than those of Comparative Example 1. Among them, the antibody activity retention rate of Example 2, which contained only S-allyl-L-cysteine, was 85.3±3.4%; the antibody protection effect of Examples 4-6, which added N-acetyl-L-histidine, was further improved, with Example 5 showing the best effect, reaching an antibody activity retention rate of 91.2±1.9%. The detection deviations of each parallel group were small, and the overall stability was excellent.

[0076] In contrast, Comparative Example 1, which uses the traditional glycerol-BSA system, showed an antibody activity retention rate of only 46.1±2.8% under the same thermal shock conditions, indicating severe damage to the antibody structure and a significant loss of activity.

[0077] The above results confirm that the protective system based on S-allyl-L-cysteine ​​in this invention can significantly improve the antibody's resistance to thermal shock and inhibit high-temperature-induced conformational denaturation and aggregation inactivation. Simultaneously, the addition of N-acetyl-L-histidine can synergistically enhance the antibody's stability under extreme temperatures. This preservation solution can meet the requirements for long-term storage at room temperature and is also suitable for short-term high-temperature scenarios such as summer transportation and on-site testing, better meeting the stringent environmental requirements of actual IVD product applications.

[0078] Application Example 3: Verification of the synergistic effect of S-allyl-L-cysteine ​​and N-acetyl-L-histidine

[0079] 1. Experimental Objective

[0080] Example 7: Based on Example 5, the S-allyl-L-cysteine ​​component was removed, and its antibody activity was compared with that of Examples 2 and 5 after storage at room temperature for 6 months. This verifies whether there is a synergistic effect between S-allyl-L-cysteine ​​and N-acetyl-L-histidine in the antibody preservation solution provided by the present invention.

[0081] 2. Experimental Materials

[0082] 1) Antibody sample: IgG antibody (purity ≥95%), concentration 1 mg / mL;

[0083] 2) Test reagents: Antibody preservation solutions provided in Examples 2, 5, and 7 of this invention, and existing conventional antibody preservation solutions (Comparative Example 1);

[0084] 3) Testing equipment: sterile centrifuge tubes, constant temperature incubator (accuracy ±0.5℃), ELISA reader, pipettes, etc.

[0085] 3. Experimental Methods

[0086] 3.1 Room temperature stability test

[0087] 3.1.1 Mix the IgG antibody with the antibody preservation solution provided in Examples 1-3 and Comparative Example 1 at a volume ratio of 1:10. After thorough mixing, dispense the mixture into sterile centrifuge tubes, seal them, and store them in a constant temperature incubator without any disturbance.

[0088] 3.1.2 Samples were taken after 6 months of storage and equilibrated at room temperature for 30 minutes. Antibody activity was detected by enzyme-linked immunosorbent assay (ELISA). Three parallel replicates were set up for each time point and each sample, and the detection was performed three times. The average antibody activity of the three detection results was calculated. The antibody activity at the initial storage time was used as the benchmark to compare and calculate the activity retention rate at that time point.

[0089] 3.2 Thermal shock resistance test

[0090] 3.2.1 Mix the IgG antibody with the antibody preservation solutions provided in Examples 2, 5, 7 and Comparative Example 1 at a volume ratio of 1:10. After thorough mixing, dispense the mixture into sterile centrifuge tubes, seal them, and place them in a constant temperature incubator at 42°C for 30 days (simulating extreme room temperature and short-term high temperature storage scenarios, which are the usual and stringent conditions for thermal shock testing). Do not disturb the tubes during this period to ensure that the thermal shock conditions are stable.

[0091] 3.2.2 After 30 days of heat shock storage, samples were taken and placed at room temperature for 30 minutes to equilibrate. Antibody activity was detected by enzyme-linked immunosorbent assay (ELISA). Each sample was tested in triplicate, and the antibody activity was calculated as the average of the three test results. The activity retention rate was calculated by comparing the antibody activity at the initial storage time.

[0092] 4. Experimental Results

[0093] 4.1 Stability test results after 6 months of storage at room temperature

[0094]

[0095] 4.2 Results of thermal shock resistance test after 30 days of thermal shock storage

[0096]

[0097] 0097.5 Result Analysis

[0098] The results of room temperature storage and high temperature thermal shock experiments show that there are significant differences in the antibody activity retention rates of Example 2 (containing only S-allyl-L-cysteine), Example 7 (containing only N-acetyl-L-histidine), and Example 5 (containing both protective components), which fully demonstrates that the two components have a significant synergistic effect.

[0099] After being stored at room temperature (25±2℃) for 6 months, the antibody activity retention rate of Example 2, which contains only S-allyl-L-cysteine, was 89.2±1.8%, while that of Example 7, which contains only N-acetyl-L-histidine, was only 78.4±1.7%. However, the antibody activity retention rate of Example 5, which contains both protective components, reached 93.3±1.7%, which was significantly higher than that of the single-component protective group.

[0100] Under the harsh conditions of 30 days of heat shock at 42°C, the above differences became even more pronounced: the antibody activity retention rate of Example 2 was 73.6±3.8%, while the antibody activity of Example 7 and Comparative Example 1 was severely lost, with only 10.3±6.7% and 9.8±8.3% remaining, respectively; while Example 5, which added two protective components at the same time, could still maintain a high antibody activity of "89.6±2.7%", showing extremely strong high-temperature stability.

[0101] The above results confirm that S-allyl-L-cysteine ​​can provide basic protection at the level of maintaining the native conformation of the antibody, while N-acetyl-L-histidine can effectively inhibit antibody molecule aggregation. The combination of the two achieves a synergistic protective effect of "1+1>2", which greatly improves the structural stability and activity retention rate of the antibody under long-term room temperature storage and extreme high temperature environment. It effectively overcomes the problem of insufficient effect of single protective agent and provides strong support for the stable application of antibody in IVD products and complex storage and transportation environments.

[0102] Application Example 4: Verification by Immunocytochemical Staining Experiment after 30 Days of Heat Shock Storage

[0103] 1. Experimental Objective

[0104] This study verifies whether the antigen-binding specificity and immunocytochemical staining effect of IgG antibodies preserved in the antibody preservation solution after being stored at 42°C for 30 days still meet the requirements of practical applications, further demonstrating the practicality of the antibody preservation solution provided by this invention and ensuring that the antibody can still be adapted to immunocytochemical detection scenarios in the IVD field after storage at extreme high temperatures.

[0105] 2. Experimental Materials

[0106] 1) Antibody samples: The antibody preservation solution of Example 5, which has been stored at 42°C for 30 days, was mixed with IgG antibody (purity ≥95%) (mixed at a volume ratio of 1:10, the same as the heat shock test sample of Example 3); at the same time, a fresh IgG antibody sample (without any storage treatment, purity ≥95%, concentration 1mg / mL) was set as a positive control, and the antibody mixture of Comparative Example 1 after 30 days of heat shock was set as a negative control.

[0107] 2) Test reagents: Immunocytochemistry kit (including secondary antibody, DAB chromogenic solution, antigen retrieval solution), PBS buffer (pH 7.4), 4% paraformaldehyde fixative, Triton X-100 permeabilization solution, blocking solution, hematoxylin staining solution, etc.

[0108] 3) Test sample: HepG2 adherent cells expressing IgG antibody-corresponding antigen were seeded on cell spread sheets and cultured to the logarithmic growth phase.

[0109] 4) Testing equipment: inverted microscope, constant temperature incubator, pipette, staining jar, cell slides, glass slides, coverslips, centrifuge, etc.

[0110] 3. Experimental Methods

[0111] 3.1 Cell sample processing: Take out the crawling slides inoculated with target cells, wash twice with PBS buffer for 3 minutes each time; add 4% paraformaldehyde fixative, fix at room temperature for 15 minutes, wash three times with PBS buffer for 5 minutes each time; add 0.3% Triton X-100 permeabilization buffer, permeabilize at room temperature for 10 minutes, wash three times with PBS buffer for 5 minutes each time to complete cell fixation and permeabilization.

[0112] 3.2 Blocking treatment: Add blocking solution to the cell slide and incubate at 37°C for 30 minutes to block non-specific binding sites. After incubation, wash three times with PBS buffer for 5 minutes each time.

[0113] 3.3 Primary antibody incubation: Add 3 antibody samples (antibody sample from Example 5 after heat shock, fresh antibody positive control, and antibody negative control from Comparative Example 1 after heat shock) to the slides, with a drop volume of 30 μL / slide. Incubate overnight at 4°C. After incubation, wash three times with PBS buffer for 5 minutes each time to remove unbound primary antibody.

[0114] 3.4 Secondary antibody incubation: Add the corresponding fluorescent secondary antibody (or enzyme-labeled secondary antibody) to all slides and incubate at 37°C for 30 minutes. After incubation, wash three times with PBS buffer for 5 minutes each time to remove unbound secondary antibody. If using fluorescent secondary antibody, wash in the dark.

[0115] 3.5. Color Development and Counterstaining: If using enzyme-labeled secondary antibody, add DAB chromogenic solution to the slide and develop the color for 2-4 minutes at room temperature in the dark. Observe the color development under an inverted microscope. After the positive cells are clearly stained, stop the color development with distilled water. Then counterstain with hematoxylin for 20 seconds, differentiate with hydrochloric acid ethanol, rinse with running water to return to blue, and wash twice with PBS buffer. If using fluorescent secondary antibody, nuclear staining can be performed directly without DAB chromogenic solution.

[0116] 3.6 Mounting and Observation: Dehydrate the slides with graded ethanol (no dehydration is required for fluorescent staining), clear them with a clearing agent, and mount them with neutral resin (use anti-fluorescence quenching mounting solution for fluorescent staining); observe the cell staining under an inverted microscope, and observe the fluorescent staining under a fluorescence microscope.

[0117] 3.7 Result Judgment: Two professional laboratory personnel independently observe the staining effect and score it from three dimensions: staining intensity, accuracy of positive localization, and background clarity (score criteria: staining intensity 0-3 points, 0 points for no staining, 3 points for strong staining; positive localization 0-2 points, 0 points for incorrect localization, 2 points for accurate localization (target antigen corresponding to cell localization); background clarity 0-2 points, 0 points for blurred background, 2 points for clean background). The total score is 7 points, ≥5 points is qualified, and ≥6 points is excellent.

[0118] 4. Experimental Results

[0119]

[0120] 5. Results Analysis

[0121] Combination Figure 1 As shown in the immunocytochemical staining results, after 30 days of heat shock storage at 42°C, the IgG antibody preserved in the antibody preservation solution of Example 5 exhibited excellent immunocytochemical staining effects (see...). Figure 1 A), with a total score of 6.4 ± 0.3, compared to the positive control (see...). Figure 1 B) No significant difference, strong staining intensity, accurate positive localization (can be clearly located to the target antigen site in the cell), clean background, can effectively identify cells expressing the target antigen, and fully meet the core requirements of immunocytochemical detection.

[0122] In contrast, the comparative antibody sample 1 after 30 days of heat shock (see Comparative Example 1) Figure 1 C), with extremely weak staining intensity, unclear positive localization, inability to accurately identify target cells, and a cluttered background, the total score was only 1.3±0.3 points, which does not meet the application requirements of immunocytochemical detection, indicating that traditional preservation solutions cannot maintain the detection performance of antibodies after storage at extreme high temperatures.

[0123] This result further verifies that in the antibody preservation solution of Example 5 of the present invention, the synergistic protective effect of S-allyl-L-cysteine ​​and N-acetyl-L-histidine not only maintains the antibody activity retention rate, but also effectively protects the antigen binding specificity of the antibody, avoids the antibody conformational abnormality caused by high temperature storage, and ensures its stable performance in actual immunocytochemical detection, effectively resisting the damage to antibody detection function caused by extreme high temperature.

[0124] In summary, the antibody preservation solution of this invention can maintain good immunoreactivity and specificity of antibodies even after storage at extreme high temperatures. It can be directly adapted to practical detection scenarios such as immunocytochemistry in the IVD field, further highlighting the practicality, innovation and application value of this invention, and providing comprehensive technical support for the long-term stable preservation of antibodies and subsequent cell-level detection applications.

[0125] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention 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 of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An antibody preservation solution, characterized in that, include: Non-animal-derived structural protectants with a mass-to-volume ratio of 0.5%-10%; An interface protectant with a mass-to-volume ratio of 0.01%-0.2%; Antioxidants with a mass-to-volume ratio of 0.01%-0.5%; A preservative with a mass-to-volume ratio of 0.01%-0.2%; 10mM-50mM buffer solution; The non-animal-derived structure protectant mentioned above includes S-allyl-L-cysteine.

2. The antibody preservation solution as described in claim 1, characterized in that, The interface protectant mentioned includes poloxamer 188.

3. The antibody preservation solution as described in claim 1, characterized in that, The antioxidants mentioned include ascorbic acid.

4. The antibody preservation solution as described in claim 1, characterized in that, The preservatives mentioned include ProClin300.

5. The antibody preservation solution as described in claim 1, characterized in that, The buffer solution is a TPS buffer system.

6. The antibody preservation solution as described in claim 1, characterized in that, Anti-aggregation synergist with a mass-to-volume ratio of 0.1%-2%.

7. The antibody preservation solution as described in claim 7, characterized in that, The anti-aggregation synergist includes N-acetyl-L-histidine.

8. A method for preserving antibodies, characterized in that, include: Add the antibody preservation solution as described in any one of claims 1-7 to the antibody sample to be preserved, and seal for preservation.

9. An antibody preservation kit, characterized in that, It includes the antibody preservation solution as described in claim 1 and the antibody preservation method as described in claim 8.

10. The use of the antibody preservation solution of claim 1, the antibody preservation method of claim 8, or the antibody preservation kit of claim 9 in antibody preservation.

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