Enzyme conjugate stabilizing solution

A technology of enzyme conjugates and stabilizing solutions, which is applied in the direction of enzyme stabilization, can solve the problems of easy loss of activity, etc., and achieve the effect of non-toxic and harmless to the human body, simple operation, and easy preparation

Active Publication Date: 2011-05-11

BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD

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AI-Extracted Technical Summary

Problems solved by technology

[0005] In view of this, the present invention proposes an enzyme conjugate stabilizing solution to solve the problem tha...

Abstract

The invention provides an enzyme conjugate stabilizing solution which comprises the following components: 1-7.5g/L protein material, 5-50g/L saccharide material, 2-30g/L amino acid material, 5-50g/L antipyrine, 1-10 g/L vitamin C, 0.2-5g/L surfactant, 0.5-5 million IU/L gentamycin sulfate, 0.5g/L Proclin 300 and 0.02mol/L phosphate buffer solution the pH value of which is 7.4. By using the enzyme conjugate stabilizing solution provided by the invention, after the enzyme conjugate is not in a low temperature environment, the bioactivity of the enzyme conjugate can be maintained for a longer time, thereby solving the problem that the enzyme conjugate is easy to lose activity after the enzyme is not in the low temperature storage environment in the related technology. In addition, the enzyme conjugate stabilizing solution provided by the invention is easy to prepare, the sensitivity is not affected, the cost is low, the operation is simple, environmental pollution can not be caused, the stabilizing solution is nontoxic and harmless to the human body and the like.

Application Domain

Enzyme stabilisation

Technology Topic

Protein proteinChemistry +10

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Examples

Experimental program(3)

Example Embodiment

[0019] Example 1

[0020] The TSH chemiluminescence immunoassay method established by preparing an anti-thyroid stimulating hormone (TSH) monoclonal antibody-horseradish peroxidase (HRP) conjugate working solution according to the embodiment of the present invention includes the following steps:

[0021] Step 1. Prepare the enzyme conjugate stabilizing solution, the components and concentrations are:

[0022]

[0023]

[0024] Step 2. Coat the white light-emitting plate with anti-TSH monoclonal antibody at a concentration of 5ug/ml, 100ul/well, and coat at 37°C for 2 hours. After washing the plate, coat with blocking solution at 200/well at 4°C for 18 hours Then, discard the sealing solution in the coating plate, pat dry, and place it in a drying oven at 37°C with a humidity of 25% for 4 hours;

[0025] Step 3. Dilute the anti-TSH monoclonal antibody-HRP enzyme conjugate with the enzyme conjugate stabilizing solution to a concentration of 1ug/ml, and then place the prepared working solution of the enzyme conjugate in a 37°C incubator for 3 days and 7 days. Take it out after 15 days, 15 days, 30 days, and 60 days, and compare with the enzyme conjugate working solution stored at 2~8℃ in parallel;

[0026] Step 4. Prepare 0μIU/ml, 0.1μIU/ml, 0.5μIU/ml, 2.5μIU/ml, 15μIU/ml, 100μIU/ml series TSH calibration products;

[0027] Step 5. Set the sample hole, add 50μl series of TSH calibration products and quality control products QCL, QCH on the TSH coating plate, then add 50μl and place in a refrigerator at 2~8℃ and a thermostat at 37℃3 The working solution of the enzyme conjugate after being destroyed in days, 7 days, 15 days, 30 days, and 60 days, after reacting at 37°C for 1 hour, wash the plate 5 times with phosphate buffered saline (PBS-T) added with Tween-20, Pat dry

[0028] Step 6. Add 100 μl of substrate solution to each well, and detect the luminescence signal value with a photon counter at 5 minutes. The results are shown in Table 1 below.

[0029] Table I

[0030]

Example Embodiment

[0031] Example 2

[0032] The TSH chemiluminescence immunoassay method established by preparing an anti-thyroid stimulating hormone (TSH) monoclonal antibody-horseradish peroxidase (HRP) conjugate working solution according to the embodiment of the present invention includes the following steps:

[0033] Step 1. Prepare the enzyme conjugate stabilizing solution, the components and concentrations are:

[0034]

[0035] Step 2. Coat the white light-emitting plate with anti-TSH monoclonal antibody at a concentration of 5ug/ml, 100ul/well, and coat at 37°C for 2 hours. After washing the plate, coat with blocking solution at 200/well at 4°C for 18 hours Then, discard the sealing solution in the coating plate, pat dry, and place it in a drying oven at 37°C with a humidity of 25% for 4 hours;

[0036] Step 3. Dilute the anti-TSH monoclonal antibody-HRP enzyme conjugate with the enzyme conjugate stabilizing solution to a concentration of 1ug/ml, and then place the prepared enzyme conjugate working solution in a 37°C incubator for 3 days and 7 days , Take out after 15 days, 30 days, and 60 days, and compare with the enzyme conjugate working solution stored at 2~8℃ in parallel;

[0037] Step 4. Prepare 0μIU/ml, 0.1μIU/ml, 0.5μIU/ml, 2.5μIU/ml, 15μIU/ml, 100μIU/ml series TSH calibration products;

[0038] Step 5. Set the sample hole, add 50μl series of TSH calibration products and quality control products QCL, QCH on the TSH coating plate, then add 50μl and place in a refrigerator at 2~8℃ and a thermostat at 37℃3 The working solution of the enzyme conjugate after being destroyed in days, 7 days, 15 days, 30 days, and 60 days, after reacting at 37°C for 1 hour, wash the plate 5 times with phosphate buffered saline (PBS-T) added with Tween-20, Pat dry

[0039] Step 6. Add 100 μl of substrate solution to each well, and detect the luminescence signal value with a photon counter at 5 minutes. The results are shown in Table 2 below.

[0040] Table II

[0041]

Example Embodiment

[0042] Example 3

[0043] The TSH chemiluminescence immunoassay method established by preparing an anti-thyroid stimulating hormone (TSH) monoclonal antibody-horseradish peroxidase (HRP) conjugate working solution according to the embodiment of the present invention includes the following steps:

[0044] Step 1. Prepare the enzyme conjugate stabilizing solution, the components and concentrations are:

[0045]

[0046] Step 2. Coat the white light-emitting plate with anti-TSH monoclonal antibody at a concentration of 5ug/ml, 100ul/well, and coat at 37°C for 2 hours. After washing the plate, coat with blocking solution at 200/well at 4°C for 18 hours Then, discard the sealing solution in the coating plate, pat dry, and place it in a drying oven at 37°C with a humidity of 25% for 4 hours;

[0047] Step 3. Dilute the anti-TSH monoclonal antibody-HRP enzyme conjugate with the enzyme conjugate stabilizing solution to a concentration of 1ug/ml, and then place the prepared enzyme conjugate working solution in a 37°C incubator for 3 days and 7 days , Take out after 15 days, 30 days, and 60 days, and compare with the enzyme conjugate working solution stored at 2~8℃ in parallel;

[0048] Step 4. Prepare 0μIU/ml, 0.1μIU/ml, 0.5μIU/ml, 2.5μIU/ml, 15μIU/ml, 100μIU/ml series TSH calibration products;

[0049] Step 5. Set the sample hole, add 50μl series of TSH calibration products and quality control products QCL, QCH on the TSH coating plate, then add 50μl and place in a refrigerator at 2~8℃ and a thermostat at 37℃3 The working solution of the enzyme conjugate after being destroyed in days, 7 days, 15 days, 30 days, and 60 days, after reacting at 37°C for 1 hour, wash the plate 5 times with phosphate buffered saline (PBS-T) added with Tween-20, Pat dry

[0050] Step 6. Add 100 μl of substrate solution to each well, and detect the luminescence signal value with a photon counter at 5 minutes. The results are shown in Table 3 below.

[0051] Table Three

[0052]

[0053] In summary, using the enzyme conjugate stabilizing solution of the present invention can keep the biological activity of the enzyme conjugate for a longer time after leaving the low-temperature environment, and can make the biological activity of the enzyme conjugate stable at 2~8℃ for up to two times. Years, 37°C stability for up to one month, which can solve the problem of the related technology that the coated plate is easy to lose activity after leaving the low-temperature storage environment. In addition, the enzyme conjugate stabilizing solution provided by the present invention is easy to prepare, does not affect sensitivity, has low cost, simple operation, no environmental pollution, and is non-toxic and harmless to the human body.

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Classification and recommendation of technical efficacy words

Easy to prepare

Enzyme conjugate stabilizing solution

A technology of enzyme conjugates and stabilizing solutions, which is applied in the direction of enzyme stabilization, can solve the problems of easy loss of activity, etc., and achieve the effect of non-toxic and harmless to the human body, simple operation, and easy preparation

AI-Extracted Technical Summary

Problems solved by technology

Abstract

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Examples

Example Embodiment

Example Embodiment

Example Embodiment

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Description & Claims & Application Information

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