CN111289757A - Production process of efficient and stable scientific research human C-reactive protein kit - Google Patents

Abstract

The invention relates to the technical field of immunology, in particular to a production process of a high-efficiency and stable scientific research human C-reactive protein kit. The method comprises the steps of preparing a coated flat-bottom microporous plate, preparing a standard mother solution, preparing a detection antibody mother solution, preparing an enzyme conjugate mother solution and preparing a diluent; wherein the preparation of the coated flat-bottom microplate comprises the following steps: (1) redissolving the coated antibody: dissolving the coated antibody by using 0.01M PBS buffer solution, and then diluting the coated antibody by using the PBS buffer solution; (2) 50 μ L of the diluted coated antibody solution was applied to an Elisa plate and left at 4 ℃ for 12 hours; taking out the coated plate, returning to room temperature, removing the coating liquid, adding the sealing liquid, shaking for 1-3 hours, and removing the sealing liquid; (3) adding a plate washing liquid, oscillating, removing the plate washing liquid, adding a coated plate protective agent, incubating for a period of time, removing the coated plate protective agent, spin-drying the liquid, vacuumizing, and sealing the plate to obtain the coated flat-bottom microporous plate.

Description

Production process of efficient and stable scientific research human C-reactive protein kit

Technical Field

The invention relates to the technical field of immunology, in particular to a production process of a high-efficiency and stable scientific research human C-reactive protein kit.

Background

C-reactive protein (CRP) is a substance which is found in 1930 by Tillett and Francis in the serum of pneumonia patients and has precipitation reaction with C-polysaccharide (CPS), a non-specific type of mycopolysaccharide component of pneumococcus (now called streptococcus pneumoniae). Macleod and Avery, in 1941, demonstrated that this protein appears during acute infection, and that the flocculent precipitation reaction with C-polysaccharide is Ca-dependent²⁺Is present. The molecular structure of CRP is a circular pentasphere composed of 5 identical, unglycosylated polypeptide subunits, each subunit consisting of a 206 amino acid, 23kDa molecular weight polypeptide chain.

The detection sensitivity of CRP is usually 3mg/L, while hypersensitive CRP can accurately give a quantitative value of 0.15-8 mg/L. CRP is synthesized and secreted by the liver, and when a body encounters stress, tissue trauma and various inflammatory stimuli, the synthesis of CRP is rapidly increased, and the concentration level reaches dozens of times or even hundreds of times of the former level. High levels of CRP can be detected within 12-48 hours after infection, and has been used as one of the indexes of inflammatory diseases in clinic, and the rising amplitude of the CRP is in positive correlation with the severity of the diseases. Conventional CRP is a sensitive indicator for observing inflammation and tissue damage, and the range of testing is typically 8-160 mg/L. The method is widely applied to infection, autoimmune diseases and the like in clinic, and is suitable for the infection of bacteria or viruses and the observation of the curative effect of antibiotics. Hypersensitivity CRP (hs-CRP), inflammation plays an important role in the formation and development process of cardiovascular and cerebrovascular and atherosclerotic plaques, and the formation and development of atherosclerotic plaques are the pathological bases of the occurrence of cardiovascular and cerebrovascular stenosis, rupture and thrombosis, and further the occurrence of myocardial infarction and cerebral thrombosis.

The stability is used as an important index for keeping the product safe and effective as an in vitro diagnostic reagent, and has important guiding significance for links such as production, transportation, storage, use and the like of the product. In-vitro diagnostic reagents are biological reagents, and some active groups have different chemical instability trends and are easy to undergo reactions such as hydrolysis, enzymolysis, oxidation and the like, so that the quality and stability of the reagents are influenced. At present, the stability of the C-reactive protein kit is mainly reflected on the reagent R2 and a calibrator, but the prior reagent R2 and the calibrator have poor stability to heat, so the storage difficulty is high, and the storage at a lower temperature is required to ensure that active ingredients in the kit do not lose efficacy.

Disclosure of Invention

Aiming at the technical problems, the invention provides a production process of a high-efficiency and stable scientific research human C-reactive protein kit, which comprises the steps of preparing a coated flat-bottom microporous plate, preparing a standard mother solution, preparing a detection antibody mother solution, preparing an enzyme conjugate mother solution and preparing a diluent; wherein the preparation of the coated flat-bottom microplate comprises the following steps:

(1) redissolving the coated antibody: dissolving the coated antibody by using 0.01M PBS buffer solution to obtain 600-700 mu g/ml coated antibody solution, and then diluting to 6-7 mu g/ml by using 0.01M PBS buffer solution;

(2) taking 50 mu L of the diluted coating antibody solution to an Elisa ELISA plate, and standing at 4 ℃ for 12 hours to obtain a coating plate; taking out the coated plate, returning to room temperature, discarding the coating solution, adding 250-350 mu L of confining liquid, oscillating at room temperature for 1-3 hours, and discarding the confining liquid;

(3) adding 250-350 mu L of plate washing liquid, shaking, removing the plate washing liquid, adding 250-350 mu L of coating plate protective agent, incubating for 20-60 min, removing the coating plate protective agent, spin-drying the liquid, vacuumizing, and sealing the plates to obtain the coated flat-bottom microporous plate.

As a preferable technical scheme, the preparation raw materials of the confining liquid in the step (2) comprise, by weight, 0.03-0.08 wt% of a nonionic surfactant, 15-25 wt% of bovine serum albumin, and the balance of a PHS buffer solution; the HLB value of the surfactant is 15-20.

As a preferred technical scheme, the nonionic surfactant is polysorbate-20 and/or polysorbate-80.

As a preferable technical scheme, the plate washing liquid in the step (3) consists of a sodium chloride aqueous solution and a nonionic surfactant, and the content of the nonionic surfactant is at least 8 wt%.

As a preferable technical scheme, the coating plate protective agent in the step (3) consists of 15-25 wt% of disaccharide and the balance of PBS buffer solution; the disaccharides include sucrose and trehalose.

As a preferred technical scheme, the weight of the sucrose and the trehalose is the same.

As a preferred technical scheme, the preparation of the standard mother solution comprises the following steps:

dissolving bovine serum albumin and trehalose which do not contain serum immunoglobulin G in a PBS buffer solution, and filtering by using a filter membrane with the aperture of 0.45 mu m to obtain a complex solution with the bovine serum albumin content of 0.5 wt% and the trehalose content of 5 wt%; then diluting the complex solution with a glycerol protective agent to obtain 10-15 ng/mL of the standard mother solution; the glycerol protective agent consists of 15-25 wt% of glycerol and the balance of the sealing liquid.

As a preferred technical scheme, the preparation of the detection antibody mother liquor comprises the following steps:

diluting the detection antibody complex solution with a glycerol protective agent to obtain 10-15 ng/mL of the standard product mother solution; the glycerol protective agent consists of 15-25 wt% of glycerol and the balance of the sealing liquid.

As a preferred technical scheme, the preparation of the enzyme conjugate mother liquor comprises the following steps:

diluting the initial solution of 3,3',5,5' -tetramethyl biphenyl diamine with an enzyme conjugate protective agent into a solution of 20-30 mu g/mL to obtain the enzyme conjugate mother solution.

As a preferred technical solution, the preparation of the diluent comprises the following steps:

0.2g KH was used₂PO₄、2.9g Na₂HPO₄·12H₂Preparing 0.01M PBS buffer solution by using O, 8g NaCl and 0.2g KCl, adding a nonionic surfactant and bovine serum albumin into the buffer solution for dissolving, and filtering by using a filter membrane with the aperture of 0.45 mu M to obtain a solution with the content of the nonionic surfactant of 0.05 wt% and the content of the bovine serum albumin of 0.5 wt%.

Detailed Description

The disclosure may be understood more readily by reference to the following detailed description of preferred embodiments of the invention and the examples included therein. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.

The terms "comprises," "comprising," "includes," "including," "has," "having," "contains," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted to include the ranges "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a range of values is described herein, unless otherwise stated, the range is intended to include the endpoints thereof and all integers and fractions within the range.

In addition, the indefinite articles "a" and "an" preceding an element or component of the invention are not intended to limit the number requirement (i.e., the number of occurrences) of the element or component. Thus, "a" or "an" should be read to include one or at least one, and the singular form of an element or component also includes the plural unless the stated number clearly indicates that the singular form is intended.

The invention provides a production process of a high-efficiency and stable scientific research human C-reactive protein kit, which comprises the steps of preparing a coated flat-bottom microporous plate, preparing a standard mother solution, preparing a detection antibody mother solution, preparing an enzyme conjugate mother solution and preparing a diluent; wherein the preparation of the coated flat-bottom microplate comprises the following steps:

(1) redissolving the coated antibody: dissolving the coated antibody by using 0.01M PBS buffer solution to obtain 600-700 mu g/ml coated antibody solution, and then diluting to 6-7 mu g/ml by using 0.01M PBS buffer solution;

(2) taking 50 mu L of the diluted coating antibody solution to an Elisa ELISA plate, and standing at 4 ℃ for 12 hours to obtain a coating plate; taking out the coated plate, returning to room temperature, discarding the coating solution, adding 250-350 mu L of confining liquid, oscillating at room temperature for 1-3 hours, and discarding the confining liquid;

(3) adding 250-350 mu L of plate washing liquid, oscillating for 1-3 hours at room temperature, removing the plate washing liquid, adding 250-350 mu L of coated plate protective agent, incubating for 20-60 min, removing the coated plate protective agent, spin-drying the liquid, and vacuumizing to seal plates to obtain the coated flat-bottom microporous plates.

In some embodiments, the coated flat-bottom microplate formulation comprises the steps of:

(1) redissolving the coated antibody: dissolving the coated antibody with 0.01M PBS buffer solution to obtain 654.5 mu g/ml coated antibody solution, and then diluting the coated antibody solution with 0.01M PBS buffer solution to 6.545 mu g/ml;

(2) taking 50 mu L of the diluted coating antibody solution to an Elisa ELISA plate, and standing overnight at 4 ℃ to obtain a coating plate; taking out the coated plate, returning to room temperature, removing the coating liquid, and patting the plate with absorbent paper until no obvious liquid exists; then adding 300 mu L of confining liquid, shaking for 2 hours at room temperature, and then removing the confining liquid;

(3) adding 300 μ L of plate washing liquid, slightly shaking at room temperature, discarding, drying on absorbent paper until no obvious liquid exists, and repeating the process for 5 times; and then adding 300 mu L of coating plate protective agent for incubation for 30min, then removing the coating plate protective agent, spin-drying liquid for 20s at a high speed by a plate spinner, vacuumizing for 20min, sealing plates with plate sealing films, then filling into a self-sealing bag, and performing sealed refrigeration at 2-8 ℃ to obtain the coated flat-bottom microporous plate.

In some embodiments, the raw materials for preparing the sealing liquid in step (2) comprise, by weight, 0.03 to 0.08 wt% of a nonionic surfactant, 15 to 25 wt% of bovine serum albumin, and the balance of a PHS buffer; the HLB value of the surfactant is 15-20.

Further, the nonionic surfactant is polysorbate-20 and/or polysorbate-80.

Further, the nonionic surfactants are polysorbate-20 and polysorbate-80, and the weight ratio of the polysorbate-20 to the polysorbate-80 is 3: 1.

further, the preparation raw materials of the blocking solution in the step (2) comprise, by weight, 0.05 wt% of a nonionic surfactant, 20 wt% of bovine serum albumin, and the balance of a PHS buffer solution.

The blocking solution is obtained by adding bovine serum albumin and a nonionic surfactant into a PBS buffer solution for dissolving, and then filtering by using a filter membrane with the aperture of 0.45 mu m.

In some embodiments, the wash liquor in step (3) consists of an aqueous sodium chloride solution and a non-ionic surfactant, the non-ionic surfactant being present in an amount of at least 8 wt%.

Further, the plate washing liquid in the step (3) consists of a sodium chloride aqueous solution and a nonionic surfactant, and the content of the nonionic surfactant is 10 wt%; the non-ionic surfactant is tween 20.

In some embodiments, the coating plate protective agent in step (3) consists of 15-25 wt% of disaccharide and the balance of PBS buffer; the disaccharides include sucrose and trehalose.

Further, the coating plate protective agent in the step (3) consists of 20 wt% of disaccharide and the balance of PBS buffer solution; the disaccharides include sucrose and trehalose.

In some embodiments, the sucrose and trehalose are the same weight. The applicant finds that the stability of the coated flat-bottom microporous plate in the kit is improved by adopting sucrose and trehalose with equal weight, so that the service life of the kit is further prolonged, and the detection accuracy is improved.

The coating Antibody component in the invention is a Mouse Anti-Human CRP capture Antibody (Mouse Anti-Human CRPCapture Antibody) purchased from R & D.

In some embodiments, the preparation of the standard master batch comprises the steps of:

dissolving bovine serum albumin and trehalose which do not contain serum immunoglobulin G in a PBS buffer solution, and filtering by using a filter membrane with the aperture of 0.45 mu m to obtain a complex solution with the bovine serum albumin content of 0.5 wt% and the trehalose content of 5 wt%; then diluting the complex solution with a glycerol protective agent to obtain 10-15 ng/mL of the standard mother solution; the glycerol protective agent consists of 15-25 wt% of glycerol and the balance of the sealing liquid.

Further, the preparation of the standard mother liquor comprises the following steps:

dissolving bovine serum albumin and trehalose which do not contain serum immunoglobulin G in a PBS buffer solution, and filtering by using a filter membrane with the aperture of 0.45 mu m to obtain a complex solution with the bovine serum albumin content of 0.5 wt% and the trehalose content of 5 wt%; then diluting the compound solution with a glycerol protective agent to obtain 12ng/mL of the standard mother solution; wherein the glycerol protectant consists of 20 wt% glycerol and the balance of the sealing liquid.

The standard of the invention comprises a Recombinant Human RCP standard (Recombinant Human CRPstrandarard) purchased from R & D.

In some embodiments, the preparation of the detection antibody stock solution comprises the following steps:

diluting the detection antibody complex solution with a glycerol protective agent to obtain 10-15 mu g/mL of detection antibody mother solution; the glycerol protective agent consists of 15-25 wt% of glycerol and the balance of the sealing liquid.

Further, the preparation of the detection antibody mother liquor comprises the following steps:

diluting the detection antibody complex solution with a glycerol protective agent to obtain 13.63 mu g/mL of detection antibody mother solution; wherein the glycerol protectant consists of 20 wt% glycerol and the balance of the sealing liquid.

The Detection Antibody component in the Detection Antibody redissolution provided by the invention is a Biotinylated Mouse Anti-Human CRP Detection Antibody (Biotinylated Mouse Anti-Human CRP Detection Antibody) purchased from R & D.

In some embodiments, the formulation of the enzyme conjugate stock solution comprises the steps of:

diluting the initial solution of 3,3',5,5' -tetramethyl biphenyl diamine with an enzyme conjugate protective agent into a solution of 20-30 mu g/mL to obtain the enzyme conjugate mother solution.

Further, the preparation of the enzyme conjugate mother liquor comprises the following steps:

diluting the initial solution of 3,3',5,5' -tetramethyl biphenyl diamine with enzyme conjugate protecting agent to 25 microgram/mL solution to obtain the enzyme conjugate mother solution. Wherein the enzyme conjugate protectant is commercially available, such as SIGMA (Cat No: 18995-250 ML-F).

In some embodiments, the formulation of the diluent comprises the steps of:

0.2g KH was used₂PO₄、2.9g Na₂HPO₄·12H₂Preparing 0.01M PBS buffer solution by using O, 8g NaCl and 0.2g KCl, adding a nonionic surfactant and bovine serum albumin into the buffer solution for dissolving, and filtering by using a filter membrane with the aperture of 0.45 mu M to obtain a solution with the content of the nonionic surfactant of 0.05 wt% and the content of the bovine serum albumin of 0.5 wt%.

Examples

Example 1: the production process of the high-efficiency and stable scientific research human C-reactive protein kit is provided, and comprises the preparation of a coated flat-bottom microporous plate, the preparation of a standard mother solution, the preparation of a detection antibody mother solution, the preparation of an enzyme conjugate mother solution and the preparation of a diluent; wherein the preparation of the coated flat-bottom microplate comprises the following steps:

(1) redissolving the coated antibody: dissolving the coated antibody with 0.01M PBS buffer solution to obtain 654.5 mu g/ml coated antibody solution, and then diluting the coated antibody solution with 0.01M PBS buffer solution to 6.545 mu g/ml;

(2) taking 50 mu L of the diluted coating antibody solution to an Elisa ELISA plate, and standing overnight at 4 ℃ to obtain a coating plate; taking out the coated plate, returning to room temperature, removing the coating liquid, and patting the plate with absorbent paper until no obvious liquid exists; then adding 300 mu L of confining liquid, shaking for 2 hours at room temperature, and then removing the confining liquid;

(3) adding 300 μ L of plate washing liquid, slightly shaking at room temperature, discarding, drying on absorbent paper until no obvious liquid exists, and repeating the process for 5 times; and then adding 300 mu L of coating plate protective agent for incubation for 30min, then removing the coating plate protective agent, spin-drying liquid for 20s at a high speed by a plate spinner, vacuumizing for 20min, sealing plates with plate sealing films, then filling into a self-sealing bag, and performing sealed refrigeration at 2-8 ℃ to obtain the coated flat-bottom microporous plate.

The preparation raw materials of the confining liquid in the step (2) comprise, by weight, 0.05 wt% of a nonionic surfactant, 20 wt% of bovine serum albumin, and the balance of a PHS buffer solution. Is prepared by dissolving bovine serum albumin and nonionic surfactant in PBS buffer solution, and filtering with filter membrane with pore diameter of 0.45 μm. The nonionic surfactant is polysorbate-20 and polysorbate-80, and the weight ratio is 3: 1.

the plate washing liquid in the step (3) consists of a sodium chloride aqueous solution and a nonionic surfactant, wherein the content of the nonionic surfactant is 10 wt%; the non-ionic surfactant is tween 20.

The coating plate protective agent in the step (3) consists of 20 wt% of disaccharide and the balance of PBS buffer solution; the disaccharide comprises sucrose and trehalose, and the weight of the sucrose and the trehalose is the same.

The preparation method of the standard mother solution comprises the following steps:

dissolving bovine serum albumin and trehalose which do not contain serum immunoglobulin G in a PBS buffer solution, and filtering by using a filter membrane with the aperture of 0.45 mu m to obtain a complex solution with the bovine serum albumin content of 0.5 wt% and the trehalose content of 5 wt%; then diluting the compound solution with a glycerol protective agent to obtain 12ng/mL of the standard mother solution; wherein the glycerol protectant consists of 20 wt% glycerol and the balance of the sealing liquid.

The preparation of the detection antibody mother liquor comprises the following steps:

diluting the detection antibody complex solution with a glycerol protective agent to obtain 13.63 mu g/mL of detection antibody mother solution; wherein the glycerol protectant consists of 20 wt% glycerol and the balance of the sealing liquid.

The preparation of the enzyme conjugate mother liquor comprises the following steps:

diluting the initial solution of 3,3',5,5' -tetramethyl diphenylenediamine (HRP initial solution, Xinaosheng (Cat No: A-2004)) with enzyme conjugate protectant to 25 μ g/mL solution to obtain the enzyme conjugate mother solution. Wherein the enzyme conjugate protectant is commercially available SIGMA (Cat No: 18995-250 ML-F).

The preparation of the diluent comprises the following steps:

0.2g KH was used₂PO₄、2.9g Na₂HPO₄·12H₂O, 8g NaCl, and 0.2g KCl 0.01M PBS bufferThen adding a nonionic surfactant and bovine serum albumin into the buffer solution for dissolving, and filtering by using a filter membrane with the aperture of 0.45 mu m to obtain a solution with the content of the nonionic surfactant of 0.05 wt% and the content of the bovine serum albumin of 0.5 wt%. The diluent was prepared into 1050. mu.L (for diluting the standard), 700. mu.L (for diluting the standard), 6mL (for diluting the detection antibody), and 12mL (for diluting the enzyme conjugate) bottles, respectively.

Then the components are assembled into a kit set for normal use, and the specific information is as follows:

TABLE 1 Standard kit Assembly

The applicant finds that in the scientific research human C-reactive protein kit in the embodiment, the reaction amplitude of the reagents of the standard substance mother liquor and the detection antibody mother liquor is not obviously changed in 7 days at 37 ℃, and the stability is good.

Example 2: the production process of the efficient and stable scientific research human C-reactive protein kit comprises the steps of preparation of a coated flat-bottom microporous plate, preparation of a standard mother solution, preparation of a detection antibody mother solution, preparation of an enzyme conjugate mother solution and preparation of a diluent. The difference from example 1 is that trehalose is not contained in the coating plate protective agent. The coated flat-bottom microporous plate in the embodiment has poor coating effect and low detection sensitivity.

Example 3: the production process of the efficient and stable scientific research human C-reactive protein kit comprises the steps of preparation of a coated flat-bottom microporous plate, preparation of a standard mother solution, preparation of a detection antibody mother solution, preparation of an enzyme conjugate mother solution and preparation of a diluent. The difference from example 1 is that the nonionic surfactant of the blocking solution is polysorbate-80. The applicant finds that in the scientific research human C-reactive protein kit in the embodiment, the concentrations of the standard mother solution and the detection antibody mother solution are generally reduced by about 35-45% in 7 days at 37 ℃, and the stability is greatly influenced.

The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in other forms, and any person skilled in the art may modify or change the technical content of the above disclosure into equivalent embodiments with equivalent changes, but all those simple modifications, equivalent changes and modifications made to the above embodiments according to the technical spirit of the present invention still belong to the protection scope of the present invention.

Claims (10)

1. A production process of a high-efficiency and stable scientific research human C-reactive protein kit is characterized by comprising the steps of preparing a coated flat-bottom microporous plate, preparing a standard mother solution, preparing a detection antibody mother solution, preparing an enzyme conjugate mother solution and preparing a diluent; wherein the preparation of the coated flat-bottom microplate comprises the following steps:

(1) redissolving the coated antibody: dissolving the coated antibody by using 0.01M PBS buffer solution to obtain 600-700 mu g/ml coated antibody solution, and then diluting to 6-7 mu g/ml by using 0.01M PBS buffer solution;

(2) taking 50 mu L of the diluted coating antibody solution to an Elisa ELISA plate, and standing at 4 ℃ for 12 hours to obtain a coating plate; taking out the coated plate, returning to room temperature, discarding the coating solution, adding 250-350 mu L of confining liquid, oscillating at room temperature for 1-3 hours, and discarding the confining liquid;

(3) adding 250-350 mu L of plate washing liquid, shaking, removing the plate washing liquid, adding 250-350 mu L of coating plate protective agent, incubating for 20-60 min, removing the coating plate protective agent, spin-drying the liquid, vacuumizing, and sealing the plates to obtain the coated flat-bottom microporous plate.

2. The process for producing the efficient and stable human C-reactive protein kit for scientific research according to claim 1, wherein the raw materials for preparing the confining liquid in the step (2) comprise, by weight, 0.03 to 0.08 wt% of a nonionic surfactant, 15 to 25 wt% of bovine serum albumin, and the balance of a PHS buffer; the HLB value of the surfactant is 15-20.

3. The process for producing the highly efficient and stable scientific human C-reactive protein kit as claimed in claim 2, wherein the non-ionic surfactant is polysorbate-20 and/or polysorbate-80.

4. The process for producing a highly efficient and stable scientific human C-reactive protein kit as claimed in claim 1, wherein the wash solution in step (3) comprises an aqueous solution of sodium chloride and a nonionic surfactant, wherein the nonionic surfactant is present in an amount of at least 8 wt%.

5. The production process of the high-efficiency and stable scientific human C-reactive protein kit as claimed in claim 1, wherein the coating plate protective agent in step (3) comprises 15-25 wt% of disaccharide and the balance of PBS buffer solution; the disaccharides include sucrose and trehalose.

6. The method of claim 5, wherein the sucrose and trehalose are in the same weight amounts.

7. The process for producing the highly efficient and stable scientific human C-reactive protein kit as claimed in claim 1, wherein the preparation of the standard mother solution comprises the steps of:

dissolving bovine serum albumin and trehalose which do not contain serum immunoglobulin G in a PBS buffer solution, and filtering by using a filter membrane with the aperture of 0.45 mu m to obtain a complex solution with the bovine serum albumin content of 0.5 wt% and the trehalose content of 5 wt%; then diluting the complex solution with a glycerol protective agent to obtain 10-15 ng/mL of the standard mother solution; the glycerol protective agent consists of 15-25 wt% of glycerol and the balance of the sealing liquid.

8. The production process of the efficient and stable scientific human C-reactive protein kit according to any one of claims 1 to 7, wherein the preparation of the detection antibody mother solution comprises the following steps:

diluting the detection antibody complex solution with a glycerol protective agent to obtain 10-15 mu g/mL of detection antibody mother solution; the glycerol protective agent consists of 15-25 wt% of glycerol and the balance of the sealing liquid.

9. The process for producing the efficient and stable scientific human C-reactive protein kit according to any one of claims 1 to 8, wherein the preparation of the enzyme conjugate mother liquor comprises the following steps:

diluting the initial solution of 3,3',5,5' -tetramethyl biphenyl diamine with an enzyme conjugate protective agent into a solution of 20-30 mu g/mL to obtain the enzyme conjugate mother solution.

10. The production process of the efficient and stable scientific human C-reactive protein kit according to any one of claims 1 to 9, wherein the preparation of the diluent comprises the following steps:

0.2g KH was used₂PO₄、2.9g Na₂HPO₄·12H₂Preparing 0.01M PBS buffer solution by using O, 8g NaCl and 0.2g KCl, adding a nonionic surfactant and bovine serum albumin into the buffer solution for dissolving, and filtering by using a filter membrane with the aperture of 0.45 mu M to obtain a solution with the content of the nonionic surfactant of 0.05 wt% and the content of the bovine serum albumin of 0.5 wt%.