CN113945711A - Processing method for reducing non-specific adsorption of magnetic beads in autoimmune antibody detection - Google Patents
Processing method for reducing non-specific adsorption of magnetic beads in autoimmune antibody detection Download PDFInfo
- Publication number
- CN113945711A CN113945711A CN202111208822.5A CN202111208822A CN113945711A CN 113945711 A CN113945711 A CN 113945711A CN 202111208822 A CN202111208822 A CN 202111208822A CN 113945711 A CN113945711 A CN 113945711A
- Authority
- CN
- China
- Prior art keywords
- working solution
- magnetic bead
- bead working
- magnetic
- calf serum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/24—Immunology or allergic disorders
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The utility model provides a processing method for reducing the nonspecific adsorption of magnetic beads in autoimmune antibody detection, which comprises the following steps: firstly, preparing a magnetic bead working solution; secondly, adding newborn calf serum into the prepared magnetic bead working solution; thirdly, uniformly mixing the magnetic bead working solution added with the newborn calf serum at room temperature, and reacting; fourthly, cleaning, standing and separating the magnetic bead working solution after the reaction with the newborn calf serum is finished, and preparing again according to the concentration of 0.3 mg/ml; fifthly, adding the magnetic bead working solution prepared after the treatment of the newborn calf serum to carry out anti-myeloperoxidase immunoglobulin G antibody detection; the treatment method is characterized in that animal serum is added into the magnetic bead working solution to combine with nonspecific substances, so that the nonspecific adsorption capacity of magnetic particles is reduced, false positive results caused by nonspecific adsorption in the detection process are obviously reduced, and the detection sensitivity and specificity are effectively improved.
Description
Technical Field
The utility model relates to the technical field of immunoassay, in particular to a processing method for reducing the nonspecific adsorption of magnetic beads in autoimmune antibody detection.
Background
Chemiluminescence immunoassay (CLIA) is started in the middle of the last 70 th century, has been developed to the present day as a mature and advanced ultra-trace active substance detection technology, and has a wide application range. The technology develops rapidly in nearly 10 years, is the fastest immunoassay method popularized and applied at present, is the most advanced labeling immunoassay technology at present, and has sensitivity and accuracy which are several orders of magnitude higher than those of an enzyme immunoassay method and a fluorescence method.
The magnetic particles are colloidal composite materials which are formed by combining magnetic nanoparticles and inorganic or organic molecules and can be uniformly dispersed in a certain base solution and have high stability. Because the magnetic particles have the characteristics of magnetic responsiveness, low cost, low energy consumption, no pollution and the like, people can fix enzymes, antibodies, oligonucleotides and other bioactive substances on the surfaces of the magnetic particles or through functional groups (such as amino, carboxyl, sulfydryl, ethylene oxide and the like) on the surfaces of the magnetic particles, and the magnetic particles can be further used in the fields of enzyme immobilization, targeted drug carriers, cell sorting, immunoassay, separation and purification of protein and nucleic acid, hybridization assay and the like. The traditional immunology detection mostly uses an enzyme label plate as a solid phase carrier, and suspended magnetic particles as the carrier have higher specific surface area, can react with a sample more fully, and have the advantages of higher sensitivity, higher detection speed, better repeatability and the like compared with the enzyme label plate carrier in addition to the flexible application of an external magnetic field, so that the traditional immunology detection is widely applied to the fields of biological and medical detection and the like.
Autoimmune diseases refer to diseases caused by the body's immune response to its antigen, which results in damage to its tissues. The common diseases include rheumatoid arthritis, systemic lupus erythematosus, sicca syndrome, scleroderma, mixed connective tissue disease, vasculitis and the like.
Non-specific adsorption of magnetic particles to non-target proteins in the serum sample can lead to false positive results, resulting in reduced overall detection sensitivity. The commonly used magnetic bead blocking agents in the prior art include BSA, serum, casein and the like, but the conventional magnetic bead blocking agents cannot meet the requirements of products, so the improvement is needed.
Disclosure of Invention
The utility model mainly aims to solve the problem of partial nonspecific adsorption of products in the prior art, and provides a treatment method for reducing the nonspecific adsorption of magnetic beads in autoimmune antibody detection, so as to achieve the technical effect of reducing the nonspecific adsorption of the surfaces of the magnetic beads to reduce the background.
In order to solve the technical problems, the utility model provides the following technical scheme:
the utility model discloses a processing method for reducing the nonspecific adsorption of magnetic beads in autoimmune antibody detection, which comprises the following steps:
the first step is as follows: preparing a magnetic bead working solution;
the second step is that: adding newborn calf serum into the prepared magnetic bead working solution;
the third step: uniformly mixing the magnetic bead working solution added with the newborn calf serum at room temperature, and reacting;
the fourth step: cleaning, standing and separating the magnetic bead working solution after the reaction with the newborn calf serum is finished, and preparing again according to the concentration of 0.3 mg/ml;
the fifth step: and adding the magnetic bead working solution prepared by processing the newborn bovine serum in the steps to detect an anti-Myeloperoxidase (MPO) IgG antibody.
The above-mentioned treatment method for reducing the non-specific adsorption of magnetic beads in the detection of autoimmune antibodies,
the magnetic bead working solution is streptavidin magnetic bead working solution with the concentration of 0.3mg/ml prepared by phosphate buffer solution;
the newborn bovine serum is special grade newborn bovine serum; the volume concentration of the newborn calf serum added into the magnetic bead working solution is 10 +/-5%;
uniformly mixing the magnetic bead working solution added with the newborn calf serum at room temperature, and reacting for 14-18 hours;
the magnetic bead working solution after the reaction with the newborn calf serum is washed for 2 times by adopting a phosphate buffer solution, the concentration of the magnetic bead working solution is kept at 0.3mg/ml during washing, and the washing time is 5-10min each time; standing and separating for at least 2min after cleaning, and then removing the separated cleaning solution;
the detection of the anti-Myeloperoxidase (MPO) immunoglobulin G (IgG) antibody is autoimmune antibody detection, and a detection test is carried out by adopting a magnetic particle chemiluminescence immunoassay method.
The treatment method for reducing non-specific adsorption in the detection of the autoimmune antibody is described above, wherein the phosphate buffer is 0.02M PBS buffer which is a common phosphate buffer.
The processing method for reducing the nonspecific adsorption of the magnetic beads in the autoimmune antibody detection has the advantages that the nonspecific adsorption of magnetic particles is reduced by adding the newborn bovine serum to the magnetic bead working solution in combination with the nonspecific substance in the serum sample, the false positive result caused by the nonspecific adsorption in the detection process is obviously reduced, and the sensitivity and specificity of the detection are effectively improved.
Detailed Description
The utility model is further illustrated with reference to specific examples. The specific embodiments described are merely illustrative of the utility model and do not limit the utility model.
The utility model discloses a processing method for reducing the nonspecific adsorption of magnetic beads in autoimmune antibody detection, which comprises the following steps:
the first step is as follows: preparing a magnetic bead working solution;
the second step is that: adding newborn calf serum into the prepared magnetic bead working solution;
the third step: uniformly mixing the magnetic bead working solution added with the newborn calf serum at room temperature, and reacting;
the fourth step: cleaning, standing and separating the magnetic bead working solution after the reaction with the newborn calf serum is finished, and preparing again according to the concentration of 0.3 mg/ml;
the fifth step: adding the magnetic bead working solution prepared after the treatment of the newborn bovine serum to carry out anti-Myeloperoxidase (MPO) immunoglobulin G (IgG) antibody detection.
The utility model relates to a processing method for reducing the nonspecific adsorption of magnetic beads in the detection of autoimmune antibodies, wherein,
the magnetic bead working solution is streptavidin magnetic bead working solution with the concentration of 0.3mg/ml prepared by phosphate buffer solution; the newborn bovine serum is special grade newborn bovine serum; the volume concentration of the newborn calf serum added into the magnetic bead working solution is 10 +/-5%; uniformly mixing the magnetic bead working solution added with the newborn calf serum at room temperature, and reacting for 14-18 hours; the magnetic bead working solution after the reaction with the newborn calf serum is washed for 2 times by adopting a phosphate buffer solution, the concentration of the magnetic bead working solution is kept at 0.3mg/ml during washing, and the washing time is 5-10min each time; standing and separating for at least 2min after cleaning, and then removing the separated cleaning solution; the detection of the anti-Myeloperoxidase (MPO) immunoglobulin G (IgG) antibody is autoimmune antibody detection, and a detection test is carried out by adopting a magnetic particle chemiluminescence immunoassay method. The phosphate buffer is commonly phosphate buffer 0.02M PBS buffer.
Example 1:
in the first step, a phosphate buffer solution is prepared by a conventional method for standby.
Secondly, preparing 10ml of streptavidin magnetic bead working solution with the concentration of 0.3mg/ml by using phosphate buffer solution, and adding 0.5ml of newborn bovine serum with the volume concentration of 5% into the prepared magnetic bead working solution.
And uniformly mixing the streptavidin magnetic bead working solution added with the newborn calf serum at room temperature, and reacting for 14 to 18 hours. Washing the reacted streptavidin magnetic bead working solution containing the newborn calf serum for 2 times by using a phosphate buffer solution, wherein the using amount of the phosphate buffer solution is 10ml in each washing, and uniformly mixing for 5-10min in each washing; standing and separating for at least 2min after cleaning, and preparing the magnetic bead working solution of 0.3mg/ml again after cleaning.
Example 2:
in the first step, a phosphate buffer solution is prepared by a conventional method for standby.
Secondly, preparing 10ml of magnetic bead working solution with the concentration of 0.3mg/ml by using phosphate buffer solution, and adding 1ml of newborn bovine serum with the volume concentration of 10 percent into the prepared magnetic bead working solution.
And uniformly mixing the streptavidin magnetic bead working solution added with the newborn calf serum at room temperature, and reacting for 14 to 18 hours. Washing the reacted streptavidin magnetic bead working solution containing the newborn calf serum for 2 times by using a phosphate buffer solution, wherein the using amount of the phosphate buffer solution is 10ml in each washing, and uniformly mixing for 5-10min in each washing; standing and separating for at least 2min after cleaning, and preparing the magnetic bead working solution of 0.3mg/ml again after cleaning.
Example 3:
in the first step, a phosphate buffer solution is prepared by a conventional method for standby.
Secondly, preparing 10ml of magnetic bead working solution with the concentration of 0.3mg/ml by using phosphate buffer solution, and adding 1.5ml of newborn bovine serum with the volume concentration of 15% into the prepared magnetic bead working solution.
And uniformly mixing the streptavidin magnetic bead working solution added with the newborn calf serum at room temperature, and reacting for 14 to 18 hours. Washing the reacted streptavidin magnetic bead working solution containing the newborn calf serum for 2 times by using a phosphate buffer solution, wherein the using amount of the phosphate buffer solution is 10ml in each washing, and uniformly mixing for 5-10min in each washing; standing and separating for at least 2min after cleaning, and preparing the magnetic bead working solution of 0.3mg/ml again after cleaning.
Example 4:
the magnetic bead working solutions prepared by the processing methods of examples 1, 2 and 3 were used to perform anti-Myeloperoxidase (MPO) immunoglobulin G (IgG) antibody detection by conventional methods, respectively, and the gradient of the luminescence value and the nonspecific size were compared.
And (3) testing the luminescence value of the sample:
in the first step, a phosphate buffer solution is prepared by a conventional method for standby.
Second, sample addition incubation: 30ul of each sample (anti-MPO-IgG antibody negative serum (N1), positive serum (P1) or positive serum (P2)) was added to the microplate, 100ul of biotin-anti-MPO antigen conjugate and 50ul of streptavidin magnetic bead working solution of newborn bovine serum prepared in example 1, 2 or 3 of the present invention were added, and the reaction solution was incubated at 37 ℃ for 30 min.
Step three, cleaning: standing the microporous plate on a magnetic plate for 5min, and pouring off supernatant in the microporous plate; then adding phosphate buffer solution into the microporous plate, uniformly mixing and cleaning; standing the microporous plate on a magnetic plate for 5min, and pouring off the phosphate buffer solution for cleaning; the washing operation was repeated 3 times, each time using 200ul of phosphate buffer.
Fourthly, incubation of enzyme-labeled secondary antibody: 100ul enzyme-labeled secondary antibody reagent is added into the micropore plate, mixed evenly and incubated for 30min at 37 ℃.
Fifthly, cleaning: standing the microporous plate on a magnetic plate for 5min, and pouring off supernatant in the microporous plate; adding cleaning buffer solution, mixing, standing the microporous plate on a magnetic plate for 5min, and pouring off the cleaning solution; the above procedure was repeated 3 times, using 200ul of washing buffer each time.
Sixth, reading: 150ul of chemiluminescent substrate was added to the plate, mixed well and then the luminescence values were measured with a chemiluminescent immunoassay analyzer, the test results are shown in Table 1.
Sample non-specific (NSB) test method:
in the first step, a phosphate buffer solution is prepared by a conventional method for standby.
Second, sample addition incubation: 30ul of each sample (anti-myeloperoxidase MPO-IgG antibody negative serum N1, positive serum P1 and P2) and 50ul of magnetic bead working solution were added to the plate, and incubated at 37 ℃ for 30 min.
Step three, cleaning: standing the microporous plate on a magnetic plate for 5min, and pouring off the supernatant in the microporous plate. Adding cleaning buffer solution, mixing, standing the microporous plate on magnetic plate for 5min, and pouring off the cleaning solution. The above procedure was repeated 3 times, using 200ul of washing buffer each time.
Fourthly, incubation of enzyme-labeled secondary antibody: 100ul enzyme-labeled secondary antibody reagent is added into the micropore plate, mixed evenly and incubated for 30min at 37 ℃.
Fifthly, cleaning: standing the microporous plate on a magnetic plate for 5min, and pouring off the supernatant in the microporous plate. Adding cleaning buffer solution, mixing, standing the microporous plate on magnetic plate for 5min, and pouring off the cleaning solution. The above procedure was repeated 3 times, using 200ul of washing buffer each time.
Sixth, reading: 150ul of chemiluminescent substrate was added to the plate, mixed well and then the luminescence values were measured with a chemiluminescent immunoassay analyzer, the test results are shown in Table 1.
Table 1: results of detection of magnetic bead working solutions prepared in examples 1, 2, and 3
| Example 1 | Example 2 | Example 3 | |
| Sample(s) | Luminous value | Luminous value | Luminous value |
| N1 | 8577 | 8319 | 9056 |
| P1 | 227604 | 265974 | 236499 |
| P2 | 946739 | 949877 | 910646 |
| NSB-N1 | 11223 | 11586 | 12730 |
| NSB-P1 | 12490 | 12863 | 12930 |
| NSB-P2 | 12782 | 12897 | 13141 |
| P1/N1 | 26.54 | 31.97 | 26.12 |
| P2/N1 | 110.38 | 114.18 | 100.56 |
Note: n1 in table: MPO-IgG antibody negative serum 1; p1: MPO-IgG antibody positive serum 1; p2: MPO-IgG antibody positive serum 2; NSB-N1: non-specific background values for serum N1; NSB-P1: non-specific background values for serum P1; NSB-P2: non-specific background values for serum P2; P1/N1: the ratio of the detection luminescence values of serum P1 and N1; P2/N1: the ratio of the luminescence values of serum P2 to N1 was determined.
Example 5:
the following substances were used in place of the newborn bovine serum added to the magnetic bead working solution prepared in example 1, respectively, to prepare a comparative example: comparative example 1 is BSA (bovine serum albumin), comparative example 2 is horse serum, and comparative example 3 is goat serum.
The luminescence value and the non-specificity were measured by a conventional test method using comparative examples No. 1, No. 2 and No. 3, respectively.
Comparative example test of luminescence value:
in the first step, a phosphate buffer solution is prepared by a conventional method for standby.
Second, sample addition incubation: 30ul of each sample (anti-MPO-IgG antibody negative serum (N1), positive serum (P1) or positive serum (P2)) was added to the microplate, 100ul of biotin-anti-MPO antigen conjugate and 50ul of streptavidin magnetic bead working solution of newborn bovine serum prepared in example 1, 2 or 3 of the present invention were added, and the reaction solution was incubated at 37 ℃ for 30 min.
Step three, cleaning: firstly, standing a microporous plate for incubating a sample on a magnetic plate for 5min, and pouring out supernatant in the microporous plate; then adding phosphate buffer solution into the microplate for incubating the samples, uniformly mixing and cleaning; standing the micro-porous plate with the incubated sample on a magnetic plate for 5min, and pouring out the phosphate buffer solution for cleaning; the washing operation was repeated 3 times, each time using 200ul of phosphate buffer.
Fourthly, incubation of enzyme-labeled secondary antibody: 100ul enzyme-labeled secondary antibody reagent is added into the micropore plate, mixed evenly and incubated for 30min at 37 ℃.
Fifthly, cleaning: standing the microporous plate on a magnetic plate for 5min, and pouring off supernatant in the microporous plate; adding cleaning buffer solution, mixing, standing the microporous plate on a magnetic plate for 5min, and pouring off the cleaning solution; the above procedure was repeated 3 times, using 200ul of washing buffer each time.
Sixth, reading: 150ul of chemiluminescent substrate was added to the microplate and mixed well, and then the luminescence value of the sample was measured and read with a chemiluminescent immunoassay analyzer, with the test results shown in Table 2.
Comparative example non-specific (NSB) test:
in the first step, a phosphate buffer solution is prepared by a conventional method for standby.
Second, sample addition incubation: 30ul of the sample (MPO-IgG negative serum N1, positive serum P1 and P2) and 50ul of the magnetic bead working solution were added to the plate, and incubated at 37 ℃ for 30 min.
And step two, cleaning: standing the microporous plate on a magnetic plate for 5min, and pouring off the supernatant in the microporous plate. Adding cleaning buffer solution, mixing, standing the microporous plate on magnetic plate for 5min, and pouring off the cleaning solution. The above procedure was repeated 3 times, using 200ul of washing buffer each time.
Thirdly, incubation of enzyme-labeled secondary antibody: 100ul enzyme-labeled secondary antibody reagent is added into the micropore plate, mixed evenly and incubated for 30min at 37 ℃.
Fourthly, cleaning: standing the microporous plate on a magnetic plate for 5min, and pouring off the supernatant in the microporous plate. Adding cleaning buffer solution, mixing, standing the microporous plate on magnetic plate for 5min, and pouring off the cleaning solution. The above procedure was repeated 3 times, using 200ul of washing buffer each time.
And fifthly, reading: 150ul of chemiluminescent substrate was added to the microplate and mixed well, and then the luminescence value of the sample was measured and read with a chemiluminescent immunoassay analyzer, with the test results shown in Table 2.
Table 2: results of detection of magnetic bead working solution prepared by treatment methods of comparative examples 1, 2 and 3
| Comparative example 1 | Comparative example 2 | Comparative example 3 | |
| Sample(s) | Luminous value | Luminous value | Luminous value |
| N1 | 11775 | 12481 | 12428 |
| P1 | 204804 | 188656 | 185644 |
| P2 | 848520 | 790519 | 781901 |
| NSB-N1 | 12475 | 13332 | 13457 |
| NSB-P1 | 13306 | 13218 | 14146 |
| NSB-P2 | 13415 | 13178 | 14301 |
| P1/N1 | 17.39 | 15.12 | 14.94 |
| P2/N1 | 72.06 | 63.34 | 62.91 |
Note: n1 in table: MPO-IgG antibody negative serum 1; p1: MPO-IgG antibody positive serum 1; p2: MPO-IgG antibody positive serum 2; NSB-N1: non-specific background values for serum N1; NSB-P1: non-specific background values for serum P1; NSB-P2: non-specific background values for serum P2; P1/N1: the ratio of the detection luminescence values of serum P1 and N1; P2/N1: the ratio of the luminescence values of serum P2 to N1 was determined.
As is clear from the data in tables 1 and 2, the luminescence values of NSB-N1, NSB-P1 and NSB-P2 in example 4 are all lower than the results in example 5, which indicates that the treatment methods in examples 1, 2 and 3 have the effect of reducing the nonspecific adsorption of magnetic beads; the ratios of the luminescence values of P1/N1 and P2/N1 in example 4 were all larger than those in example 5, indicating that the treatment methods in examples 1, 2 and 3 have the effect of improving the detection sensitivity. Therefore, the treatment method for reducing the nonspecific adsorption of the magnetic beads in the detection of the autoimmune antibody has obvious effect and effectively improves the sensitivity of the detection reagent.
The content that will not be described in this embodiment is the prior art, and therefore, the description thereof is omitted.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and all simple modifications, equivalent variations and modifications made to the above embodiment according to the technical spirit of the present invention still fall within the scope of the technical solution of the present invention.
Claims (3)
1. A processing method for reducing the nonspecific adsorption of magnetic beads in the detection of autoimmune antibodies is characterized by comprising the following steps:
the first step is as follows: preparing a magnetic bead working solution;
the second step is that: adding newborn calf serum into the prepared magnetic bead working solution;
the third step: uniformly mixing the magnetic bead working solution added with the newborn calf serum at room temperature, and reacting;
the fourth step: cleaning, standing and separating the magnetic bead working solution after the reaction with the newborn calf serum is finished, and preparing again according to the concentration of 0.3 mg/ml;
the fifth step: and adding the magnetic bead working solution prepared after the treatment of the newborn calf serum into the steps to perform anti-myeloperoxidase immunoglobulin G antibody detection.
2. The method of claim 1, wherein the magnetic beads are non-specifically adsorbed on the surface of the sample,
the magnetic bead working solution is streptavidin magnetic bead working solution with the concentration of 0.3mg/ml prepared by phosphate buffer solution;
the newborn bovine serum is special grade newborn bovine serum; the volume concentration of the newborn calf serum added into the magnetic bead working solution is 10 +/-5%;
uniformly mixing the magnetic bead working solution added with the newborn calf serum at room temperature, and reacting for 14-18 hours;
the magnetic bead working solution after the reaction with the newborn calf serum is washed for 2 times by adopting a phosphate buffer solution, the concentration of the magnetic bead working solution is kept at 0.3mg/ml during washing, and the washing time is 5-10min each time; standing and separating for at least 2min after cleaning, and then removing the separated cleaning solution;
the detection of the anti-myeloperoxidase immunoglobulin G antibody is autoimmune antibody detection, and is a detection test by adopting a magnetic particle chemiluminescence immunoassay method.
3. The method of claim 2, wherein the magnetic beads are non-specifically adsorbed on the surface of the sample,
the phosphate buffer is commonly phosphate buffer 0.02M PBS buffer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111208822.5A CN113945711A (en) | 2021-10-18 | 2021-10-18 | Processing method for reducing non-specific adsorption of magnetic beads in autoimmune antibody detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111208822.5A CN113945711A (en) | 2021-10-18 | 2021-10-18 | Processing method for reducing non-specific adsorption of magnetic beads in autoimmune antibody detection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113945711A true CN113945711A (en) | 2022-01-18 |
Family
ID=79330935
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111208822.5A Withdrawn CN113945711A (en) | 2021-10-18 | 2021-10-18 | Processing method for reducing non-specific adsorption of magnetic beads in autoimmune antibody detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113945711A (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103033624A (en) * | 2012-12-16 | 2013-04-10 | 天津市协和医药科技集团有限公司 | Human myeloperoxidase chemiluminescent immunodetection kit |
| CN103091306A (en) * | 2013-01-23 | 2013-05-08 | 苏州浩欧博生物医药有限公司 | Solution for sealing and preserving magnetic nanoparticles |
| CN105954266A (en) * | 2016-04-20 | 2016-09-21 | 北京中航赛维生物科技有限公司 | Magnetic particle-based quantitative chemiluminescent assay kit for anti-ribosome P protein antibody IgG, and preparation and detection methods thereof |
| US20170305983A1 (en) * | 2015-02-10 | 2017-10-26 | Shenzhen New Industries Biomedical Engineering Co., Ltd | Cardiac troponin i ultra-sensitive detection reagent kit, and ultra-sensitive detection method therefor |
| US20170363627A1 (en) * | 2015-02-10 | 2017-12-21 | Shenzhen New Industries Biomedical Engineering Co., Ltd. | Reagent kit used for detecting lipoprotein-associated phospholipase a2, and preparation method and application for reagent kit |
| CN107966563A (en) * | 2016-06-30 | 2018-04-27 | 深圳市亚辉龙生物科技股份有限公司 | A kind of antimyeloperoxidase antibody IgG chemiluminescence immunoassay kits and preparation method thereof |
| CN111521777A (en) * | 2020-04-30 | 2020-08-11 | 四川携光生物技术有限公司 | Treatment method for reducing non-specific adsorption in autoimmune antibody detection and kit thereof |
-
2021
- 2021-10-18 CN CN202111208822.5A patent/CN113945711A/en not_active Withdrawn
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103033624A (en) * | 2012-12-16 | 2013-04-10 | 天津市协和医药科技集团有限公司 | Human myeloperoxidase chemiluminescent immunodetection kit |
| CN103091306A (en) * | 2013-01-23 | 2013-05-08 | 苏州浩欧博生物医药有限公司 | Solution for sealing and preserving magnetic nanoparticles |
| US20170305983A1 (en) * | 2015-02-10 | 2017-10-26 | Shenzhen New Industries Biomedical Engineering Co., Ltd | Cardiac troponin i ultra-sensitive detection reagent kit, and ultra-sensitive detection method therefor |
| US20170363627A1 (en) * | 2015-02-10 | 2017-12-21 | Shenzhen New Industries Biomedical Engineering Co., Ltd. | Reagent kit used for detecting lipoprotein-associated phospholipase a2, and preparation method and application for reagent kit |
| CN105954266A (en) * | 2016-04-20 | 2016-09-21 | 北京中航赛维生物科技有限公司 | Magnetic particle-based quantitative chemiluminescent assay kit for anti-ribosome P protein antibody IgG, and preparation and detection methods thereof |
| CN107966563A (en) * | 2016-06-30 | 2018-04-27 | 深圳市亚辉龙生物科技股份有限公司 | A kind of antimyeloperoxidase antibody IgG chemiluminescence immunoassay kits and preparation method thereof |
| CN111521777A (en) * | 2020-04-30 | 2020-08-11 | 四川携光生物技术有限公司 | Treatment method for reducing non-specific adsorption in autoimmune antibody detection and kit thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108982834B (en) | Method for detecting biological molecules by using nano enzyme immune sandwich novel technology | |
| EP2817423B1 (en) | Methods and systems for signal amplification of bioassays | |
| CN112285353A (en) | Method for improving anti-biotin interference capability and sensitivity of chemiluminescence kit of streptavidin-biotin reaction system | |
| CN113049811A (en) | Nano magnetic bead coating material, preparation method thereof, detection reagent and detection kit | |
| EP0202688B1 (en) | Method and composition for detecting analyte moieties | |
| CN110988325B (en) | Blocking agent and kit containing same | |
| CN114252591A (en) | Magnetic bead coating and preparation method thereof and detection kit | |
| CN111983221B (en) | Surface-modified magnetic bead and preparation method and application thereof | |
| CN107290518B (en) | A kind of reagent for eliminating immune response false positive | |
| CN110967493A (en) | Vitamin B12 determination kit and preparation method thereof | |
| CN113945711A (en) | Processing method for reducing non-specific adsorption of magnetic beads in autoimmune antibody detection | |
| CN111693721A (en) | Preparation method and application of enzyme-linked immunosorbent assay based on prussian blue nano enzyme label | |
| CN108359642B (en) | Hybridoma cell of shrimp tropomyosin monoclonal antibody and application thereof | |
| CN112180102B (en) | Peanut allergen specificity IgE quantitative detection reagent, kit and application thereof | |
| CN108303543B (en) | Swine fever E2 protein antibody detection kit and detection method thereof | |
| CN116381222A (en) | Serotonin luminescent immune detection method and serotonin detection kit | |
| CN110878296B (en) | High-sensitivity HRP enzyme and preparation method and application thereof | |
| CN112129933B (en) | Reagent, kit and method for resisting biological interference in immunoassay system | |
| CN1250969C (en) | Test device and method for making qualitative and/or quantitative analysis to object | |
| CN117405470A (en) | Immunomagnetic bead kit and application method thereof | |
| CN111289759B (en) | SAA detection kit and SAA quantitative detection method | |
| CN113866157A (en) | Application of polyethyleneimine compound in luminescent method in-vitro diagnostic reagent | |
| CN110887968A (en) | Quantitative detection method of alpha-fetoprotein | |
| CN116840489A (en) | Detection kit for house dust mite allergen specific IgE antibody and application thereof | |
| JP3717735B2 (en) | Solid phase carrier for immunosolid phase assay |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20220118 |
|
| WW01 | Invention patent application withdrawn after publication |