CN114874994A - Hybridoma cell strain monoclonal antibody capable of secreting anti-human IgG3 monoclonal antibody and application - Google Patents
Hybridoma cell strain monoclonal antibody capable of secreting anti-human IgG3 monoclonal antibody and application Download PDFInfo
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- C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
- C07K16/4283—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an allotypic or isotypic determinant on Ig
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Abstract
The invention provides a hybridoma cell strain secreting IgG1 monoclonal antibody of anti-human IgG3 antigen, which is preserved in China Center for Type Culture Collection (CCTCC) at 11 months and 10 days 2021, and the preservation number is CCTCC NO: c2021101, named hybridoma cell line 7C 11. The invention also provides a monoclonal antibody secreted and generated by the hybridoma cell strain secreting the IgG1 monoclonal antibody of the anti-human IgG3 antigen and application thereof. A hybridoma cell strain 7C11 capable of being stably passaged and secreting an anti-human IgG3 monoclonal antibody is obtained by immunizing a BALB/C mouse by using human IgG as an antigen, fusing cells, screening and cloning, the anti-human IgG3 monoclonal antibody secreted by the cell strain can perform a specific reaction with human IgG3, the monoclonal antibody is an anti-human IgG3 monoclonal antibody, an IgG3 type anti-s sensitized cell is used for performing a test tube agglutination test to detect that the titer of the anti-human IgG3 monoclonal antibody is 1: 256, and the titer of the anti-human IgG3 monoclonal antibody is 1: 50000 measured by an ELISA method.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a hybridoma cell strain secreting an anti-human IgG3 antigen, a monoclonal antibody and application thereof.
Background
IgG is the main component of immunoglobulin in human serum, and accounts for 75% of the total amount of immunoglobulin in serum. The main function of IgG is to participate in immune regulation, and IgG can be divided into four subclasses according to the difference of the antigenicity of the γ chain in IgG molecule: IgG1, IgG2, IgG3 and IgG4 have relationship with the severity of diseases, IgG1 and IgG3 are serious pathogenicity, and the two subclasses play an important role in activating complement by combining with Fc receptors of natural killer cells and mononuclear macrophages, so that clinical detection of the IgG subclass level has auxiliary diagnosis value on neonatal hemolysis and autoimmune hemolysis diseases.
IgG is the only antibody capable of passing through the maternal placental barrier, the level of IgG subclasses in the maternal host is closely related to the occurrence of fetal neonatal hemolysis, IgG1 and IgG3 are more likely to cause hemolysis than IgG2 and IgG4, so the detection of the levels of maternal IgG1 and IgG3 can assist in the clinical diagnosis of fetal or neonatal hemolysis.
The erythrocyte autoantibodies are the causative factors of autoimmune hemolytic anemia, IgG, IgM and IgA are 3 common anti-erythrocyte autoantibody subtypes, IgG is the most common, and IgG subclass influences the degree of shortening of erythrocyte life span, compared with IgG2 and IgG4, IgG1 and IgG3 obviously shorten half-life period of erythrocytes, and the autoantibodies in IgG class are mainly directed against epitope of Rh system, so that detection of IgG1 and IgG3 level can assist clinical diagnosis of autoimmune hemolytic anemia caused by the erythrocyte autoantibodies in IgG class.
Disclosure of Invention
In order to detect human IgG3 antigen, the invention aims to provide a hybridoma cell strain secreting IgG1 monoclonal antibody of anti-human IgG3 antigen, the invention uses human IgG as antigen to prepare a hybridoma cell secreting IgG1 monoclonal antibody of anti-human IgG3 antigen by a hybridoma technology, and cell culture supernatant is concentrated to be used as human IgG3 detection reagent to be applied to detection of human IgG 3.
The invention aims to provide a hybridoma cell strain secreting IgG1 monoclonal antibody of anti-human IgG3 antigen, which is named as: hybridoma cell line 7C 11: deposited in China center for type culture Collection, address: wuhan university in Wuhan, China, with a preservation date of 2021, 11 months and 10 days and a preservation number of CCTCC NO: C2021101.
the second purpose of the invention is to provide an anti-human IgG3 monoclonal antibody, wherein the anti-human IgG3 monoclonal antibody is secreted and produced by the hybridoma cell strain secreting IgG1 monoclonal antibody of anti-human IgG3 antigen.
The type of the anti-human IgG3 monoclonal antibody is IgG1 type; the anti-human IgG3 monoclonal antibody specifically reacts with human IgG 3.
The third purpose of the invention is to provide a human IgG3 detection reagent, which comprises the anti-human IgG3 monoclonal antibody.
The fourth purpose of the invention is to provide a human IgG3 detection device, which comprises the anti-human IgG3 monoclonal antibody.
The fifth purpose of the invention is to provide a method for detecting human IgG3, which is characterized by comprising the step of contacting a sample to be detected with the anti-human IgG3 monoclonal antibody to determine whether the sample to be detected contains human IgG 3.
The sixth purpose of the invention is to provide the application of the anti-human IgG3 monoclonal antibody in the detection of human IgG 3.
The invention has the advantages that: discloses a hybridoma cell strain secreting IgG1 monoclonal antibody of anti-human IgG3 antigen and application of the monoclonal antibody; a hybridoma cell strain 7C11 capable of stably passaging and secreting an anti-human IgG3 monoclonal antibody is obtained by immunizing a BALB/C mouse by using human IgG as an antigen, and performing cell fusion, screening and cloning; the type of the anti-human IgG3 monoclonal antibody secreted by the hybridoma cell strain is IgG 1; the anti-human IgG3 monoclonal antibody can specifically react with human IgG 3; collecting the cell culture supernatant, concentrating by 10 times, and detecting with IgG3 type anti-s sensitized cell by test tube agglutination test to obtain titer of 1: 256; the antibody titer can reach 1: 50000 by the concentrated solution ELISA method.
The above description is only an overview of the technical solutions of the present invention, and in order to make the technical means of the present invention more clear and clear, and to implement the technical means according to the content of the description, some examples are described in detail below. Specific embodiments of the present invention are given in detail by the following examples.
Drawings
FIG. 1 is a photograph showing the results of ELISA-based potency assay of anti-human IgG3 monoclonal antibody;
FIG. 2 is a graph showing the results of the assay of the agglutination potency of the anti-human IgG3 monoclonal antibody in a test tube;
FIG. 3 is a picture of the result of identifying the agglutination specificity of the anti-human IgG3 monoclonal antibody in vitro;
FIG. 4 is a photograph showing the results of Western blotting to identify the anti-human IgG3 monoclonal antibody;
FIG. 5 is a photograph showing the result of identifying the subclass of anti-human IgG3 monoclonal antibody Ig;
FIG. 6 is a photograph showing the result of karyotype analysis of hybridoma cell line 7C 11.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. 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.
Example 1:
obtaining hybridoma cells;
1. preparation of immunogens
Diluting the third generation immunoglobulin (Wuhan blood products Co., Ltd. of the national drug group) with normal saline to 1mg/mL, adding aluminum phosphate adjuvant at a ratio of 9: 1, and mixing uniformly for later use, wherein the immunogen needs to be prepared just before use;
2. animal immunization and blood test
(1) Immunizing BALB/c mice with 4 weeks of age and weight of about 20g with immunogen at a dose of 100 mug/mouse, and performing subcutaneous multi-point injection;
(2) immunizing once every other week;
(3) after three times of immunization, tail vein boosting immunization is carried out, the antigen dose is 100 mug/mouse, 100 muL/mouse is collected in orbital veins after 3 days, centrifugation is carried out for 15min at 3500rpm, serum is separated, and serum titer detection is carried out; test tube agglutination titer; measuring the titer of the mouse serum antibody by an ELISA method to be more than 1: 10000 to carry out a cell fusion test;
3. cell fusion
(1) 5-7 days before fusion, performing expanded culture on NS-1 cells, and adjusting cell state to reach logarithmic growth phase with cell survival rate of above 95%; taking NS-1 cells in logarithmic phase out of the incubator, gently shaking the cells, collecting cell suspension in a 50mL centrifuge tube, centrifuging at 1000rpm for 10min, discarding supernatant, and re-suspending the cells with 10mL 1640 culture solution for later use;
(2) extracting BALB/c mouse spleen cells qualified in blood test, uniformly mixing NS-1 cells and the mouse spleen cells according to the proportion of 1: 5-1: 8, centrifuging at 1000rpm for 15min, and removing supernatant;
(3) fusing for 2min by using 50% of PEG4000 as a fusing agent, adding 50mL of 1640 culture solution to terminate the reaction, centrifuging at 800rpm for 5min, and removing the supernatant;
(4) resuspending the cell precipitate with HAT culture solution, uniformly adding the cells into a 96-well culture plate, and culturing in a 5% CO2 incubator at 37 deg.C;
screening and cloning of hybridoma cells
(1) Enzyme label plate coating and sealing
Firstly, preparing 50 mu g/ml of Human IgG1 (manufacturer: Fitzgerald), Human IgG2 (manufacturer: Fitzgerald), Human IgG3 (manufacturer: Fitzgerald) and Human IgG4 (manufacturer: Abcam) by using a coating solution, adding the mixture into an enzyme label plate according to 100 mu L/hole, and incubating for 2 hours at 37 ℃;
secondly, after coating is finished, lightly knocking off liquid in the plate, adding 200 mu L of 2% BSA (factory: SIGMA) solution into each hole, and incubating for 2 hours at 37 ℃; after the sealing is finished, washing the plate by using washing liquor (PBST), washing each plate for 5 times, and slightly knocking off residual liquid in the plate;
(2) cell selection and cloning
Culturing the fused cells in an incubator for 5-7 days, and cloning about hundreds of cells for screening;
and secondly, taking the supernatant of the hybridoma cells to a 96-well plate, wherein the volume of the supernatant is 180 mu l/well. And respectively adding 40 mul of samples to be detected into 40 mul/hole of the ELISA plates coated by the four IgG antigens, and incubating for 1 hour at 37 ℃. After the plate is washed by washing liquor (PBST), each plate is washed for 5 times, and residual liquid in the plate is slightly knocked off;
③ diluting HRP-labeled goat anti-mouse IgG (Beijing Ding Guoshang biotechnology, Inc.) with washing liquor (PBST) at a ratio of 1: 10000, adding 100 mu l/hole into an ELISA plate, and incubating for 1 hour at 37 ℃; after the plate is washed by using washing liquor (PBST), each plate is washed for 5 times, and residual liquid in the plate is slightly knocked off;
adding 100 mul of TMB color development liquid (solemn union organisms) into each hole, and adding 100 mul of stop liquid (2M H) into each hole after the color development is about 5min 2 SO 4 ) (ii) a Detecting the OD value of each hole by using a microplate reader (the wavelength is 450 nm);
culturing and cloning IgG3 specific positive hole cells to obtain a hybridoma cell strain 7C11 capable of stably secreting anti-human IgG3 monoclonal antibody; after multiple passages and multiple freeze thawing recovery, the cell strain can well grow and can stably secrete antibodies. After the expanded culture, the cells were cryopreserved and stored in liquid nitrogen.
Example 2:
preparation of anti-human IgG3 monoclonal antibody
1. The cell line 7C11 obtained in example 1 was subjected to scale-up culture using 10% complete 1640 medium;
2. collecting cell culture supernatant, concentrating the cell culture supernatant by 10 times by using a hollow fiber column, sterilizing to obtain an anti-human IgG3 monoclonal antibody concentrated solution, and measuring the antibody titer by an ELISA method to reach 1: 50000;
3. according to production requirements, the concentrated anti-human IgG3 monoclonal antibody is properly diluted, and an antibody stabilizing agent is added according to requirements, so that the concentrated anti-human IgG3 monoclonal antibody can be used as an anti-human IgG3 monoclonal antibody reagent for identifying human IgG3 or used as a raw material of a kit related to human IgG subclass detection.
Example 3:
anti-human IgG3 monoclonal antibody titer determination
1. ELISA method for measuring titer of anti-human IgG3 monoclonal antibody
(1) Coating quilt
Preparing Human IgG3 (manufacturer: Fitzgerald) into 50 mug/ml by using coating solution, adding 100 mug L/hole into an enzyme label plate, and incubating for 2 hours at 37 ℃;
(2) sealing of
After coating, slightly knocking off liquid in the plate, adding 200 mu L of 2% BSA (manufacturer: SIGMA) solution into each hole, incubating for 2 hours at 37 ℃, washing the plate by PBST after closing, washing each plate for 5 times, and slightly knocking off residual liquid in the plate;
(3) incubation of anti-human IgG3 monoclonal antibody prepared in example 2
Adding 100 mu l/hole of the anti-human IgG3 monoclonal antibody prepared in example 2 into an enzyme label plate, incubating for 1 hour at 37 ℃, washing the plate with a washing solution (PBST) 5 times per plate after the incubation is finished, and slightly knocking off residual liquid in the plate;
(4) incubation secondary antibody
HRP-labeled goat anti-mouse IgG (Beijing ancient Changsheng biotechnology, LLC) was diluted 1: 10000 with wash solution (PBST), 100. mu.l/well was added to the microplate, and incubated at 37 ℃ for 1 hour. After the end, the plates are washed by PBST, each plate is washed for 5 times, and residual liquid in the plates is slightly knocked off;
(5) color development
Adding 100 mul of TMB color development liquid (solemn union organisms) into each hole, and adding 100 mul of stop solution (2M H) into each hole after the color development is about 5min 2 SO 4 ) (ii) a Detecting the OD value of each hole by using a microplate reader (the wavelength is 450 nm);
as shown in FIG. 1, the titer of the anti-human IgG3 monoclonal antibody prepared in example 2 was 1: 50000 measured by ELISA;
2. determination of anti-human IgG3 monoclonal antibody potency by saline test tube method
(1) Preparation of anti-s sensitized cells: putting 200 mul of an anti-s antibody (IgG 3 type, manufacturer: DIAGAST) into a clean test tube, adding 200 mul of s-positive packed red blood cells of a healthy blood donor, uniformly mixing, incubating for 30min at 37 ℃, washing for 3 times by using normal saline, and preparing into an anti-s sensitized cell suspension with the concentration of 2% by using the normal saline for later use;
(2) taking a column of test tubes, adding 200 ul of physiological saline into each tube, adding 200 ul of anti-human IgG3 monoclonal antibody concentrated solution into the No. 2 tube, sequentially diluting by a pipettor by a multiple ratio, and removing 200 ul from the last tube after dilution;
(3) 200 μ l of 2% anti-s-sensitized cell suspension was added to each tube. Incubate at room temperature for 5min and immediately centrifuge at 1000rpm for 1 min. Taking out the test tube, observing and recording the result;
(4) the control should not agglutinate, and the maximum dilution of the antibody to be detected still can cause the agglutination of the erythrocytes visible by naked eyes to be used as the titer of the antibody;
as shown in FIG. 2, the test tube agglutination titer of the anti-human IgG3 monoclonal antibody prepared in example 2 was 1: 256.
Example 4:
specific identification of anti-human IgG3 monoclonal antibody
1. ELISA method for identifying specificity of anti-human IgG3 monoclonal antibody
(1) Coating quilt
Preparing 50 mu g/ml of Human IgG1 (manufacturer: Fitzgerald), Human IgG2 (manufacturer: Fitzgerald), Human IgG3 (manufacturer: Fitzgerald) and Human IgG4 (manufacturer: Abcam) by using a coating solution, adding the mixture into an ELISA plate according to 100 mu L/hole, and incubating for 2 hours at 37 ℃;
(2) sealing of
After coating, slightly knocking off the liquid in the plate, adding 200 mu L of 2% BSA (manufacturer: SIGMA) solution into each hole, and incubating for 2 hours at 37 ℃; after the sealing is finished, washing the plate by using washing liquor (PBST), washing each plate for 5 times, and slightly knocking off residual liquid in the plate;
(3) incubation of anti-human IgG3 monoclonal antibody prepared in example 2
Adding 100 ul of the anti-human IgG3 monoclonal antibody prepared in the example 2 into an enzyme labeled plate, incubating for 1 hour at 37 ℃, washing the plate with PBST after the incubation is finished, washing each plate for 5 times, and slightly knocking off residual liquid in the plate;
(6) incubation secondary antibody
HRP-labeled goat anti-mouse IgG (Beijing ancient Changsheng biotechnology, LLC) was diluted 1: 10000 with wash solution (PBST), 100. mu.l/well was added to the microplate, and incubated at 37 ℃ for 1 hour. After the plate is washed by washing liquor (PBST), each plate is washed for 5 times, and residual liquid in the plate is slightly knocked off;
(7) color development
Adding 100 mul of TMB color development liquid (solemn union organisms) into each hole, and adding 100 mul of stop solution (2M H) into each hole after the color development is about 5min 2 SO 4 ). Detecting the OD value of each hole by using a microplate reader (the wavelength is 450 nm);
the results are shown in Table 1, and the results of measuring the specificity of the anti-human IgG3 monoclonal antibody prepared in example 2 by ELISA method:
TABLE 1 result of ELISA method specific identification of anti-human IgG3 monoclonal antibody
| Antigen type | IgG1 | IgG2 | IgG3 | IgG4 |
| Absorbance value | 0.035 | 0.014 | 1.475 | 0.068 |
And (4) conclusion: the anti-human IgG3 monoclonal antibody reacts with human IgG3 antigen, does not react with human IgG1, IgG2 and IgG4 antigen, and has qualified specificity identification;
2. identification of anti-human IgG3 monoclonal antibody specificity by saline test tube method
(1) Preparation of sensitized cells: respectively taking 200 mul of anti-D antibody (IgG 1 type, manufacturer: Millipore), anti-E antibody (IgG 1 type, manufacturer: Millipore), anti-Fya antibody (IgG 1 type, manufacturer: DIAGAST), anti-S antibody (IgG 3 type, manufacturer: DIAGAST), anti-S antibody (IgG 3 type, manufacturer: DIAGAST) in a clean test tube, respectively adding 200 mul of corresponding antigen positive packed red blood cells of a healthy donor, uniformly mixing, incubating for 30min at 37 ℃, and washing for 3 times by physiological saline;
(2) taking anti-D, anti-E, anti-Fya, anti-S, anti-S sensitized cells and 3 human A 1 B, O type erythrocyte is washed by normal saline for 3 times, centrifuged at 2000rpm for 5min, supernatant is removed, and normal saline is used for preparing 2 percent erythrocyte suspension;
(3) arranging the test tubes into 2 rows, marking, adding 100 mu l of samples to be detected into the first row of test tubes, and then adding 2% of anti-D, anti-E, anti-Fya, anti-S, anti-S sensitized cells and 3 persons A into each test tube respectively 1 100 mul of B, O type red blood cell suspension; the second row of test tubes is used as erythrocyte suspension control, namely, a sample to be detected is changed into a physiological sodium chloride solution, the test tubes are placed at room temperature for reaction for 15 minutes, centrifuged at 1000rpm for 1 minute, the test tubes are shaken lightly, and whether agglutination exists or not is observed by naked eyes;
(4) the agglutination of the anti-human IgG3 monoclonal antibody prepared in example 2 with the corresponding erythrocytes was observed and recorded under the precondition that no agglutination was observed in the control group of erythrocyte suspension. Otherwise, if the control group has agglutination reaction, the test is not established, and the test needs to be repeated;
as shown in FIG. 3, the monoclonal antibody against human IgG3 agglutinated with the anti-S and anti-S sensitized cells, and with the anti-D, anti-E, anti-Fya sensitized cells and A 1 、B、O cells do not have agglutination reaction, do not have hemolysis and other phenomena which are not easy to distinguish;
3. identification of anti-human IgG3 monoclonal antibody specificity by Western blot assay
(1) Gel formulation
Preparing 5% of concentrated glue and 10% of separation glue;
(2) sample processing
Respectively taking 5 mu g of Human IgG3 (manufacturer: Fitzgerald) in 2 tubes of 1.5mL EP, adding 5 mu L of reducing loading buffer into one tube, adding 5 mu L of non-reducing loading buffer into the other tube, and heating for 10min at 100 ℃;
(3) SDS-PAGE electrophoresis
And (4) electrophoretic sample loading, wherein the treated Human IgG3 is loaded respectively, and the loading amount of the protein Marker is 5 mu L. Performing constant-voltage electrophoresis, wherein the initial voltage is 80V, the voltage is adjusted to 120V when the gel enters the separation gel, and the electrophoresis is stopped when bromophenol blue migrates to the gel bottom;
(4) rotary film
After the electrophoresis is finished, taking out the gel, cutting the gel according to the size of the protein, and shearing the PVDF membrane according to the size of the gel; the PVDF membrane is soaked in methanol for 2min and then is washed twice by adding purified water. Placing the filter paper, the gel, the PVDF membrane and the filter paper in a transfer tank in sequence, and transferring the membrane for 1h at a constant current of 200 mA;
(5) sealing of
After the membrane transfer, the PVDF membrane is placed in a pre-prepared 5% BSA (PBST prepared) blocking solution and blocked for 1h at 37 ℃. After the sealing is finished, washing for 5 times by PBST, 5min each time;
(6) incubation of anti-human IgG3 monoclonal antibodies
Placing the PDF membrane in the anti-human IgG3 monoclonal antibody prepared in example 2, incubating for 1h at 37 ℃, washing for 5 times with PBST after the incubation is finished, and each time for 5 min;
(7) incubation secondary antibody
10mL of HRP-labeled goat anti-mouse IgG (diluted 1: 3000) was added and incubated at 37 ℃ for 1 h. Washing with PBST for 5 times, each for 5 min;
(8) color development
The color development was carried out using a DAB horseradish peroxidase color development kit (manufacturer: Biyun Tian) according to the instructions.
The results are shown in fig. 4, lane 1 is human IgG3 reduced electrophoresis, lane 2 is human IgG3 non-reduced electrophoresis, anti-human IgG3 monoclonal antibody reacts with human IgG3 and strongly reacts with the heavy chain of human IgG3, and thus it can be seen that anti-human IgG3 monoclonal antibody can specifically bind to human IgG3, and thus the antibody is identified as anti-human IgG3 monoclonal antibody.
Example 5:
mouse monoclonal antibody Ig subclass identification
The IgG3 monoclonal antibody Ig subclass of example 2 was identified using a commercial kit (mouse monoclonal antibody Ig class/subclass/identification ELISA kit, manufacturer: Yikang organism);
as shown in FIG. 5, the anti-human IgG3 monoclonal antibody was IgG1 type.
Example 6:
chromosome karyotyping of hybridoma cell lines
1. After subculturing the hybridoma cell line 7C11 obtained in example 1 for 48 hours, colchicine was added to make the final concentration of colchicine 0.8. mu.g/mL, and the culture was continued at 37 ℃ for 4-6 hours. Centrifuging the above cells at 1000rpm for 10min, and removing supernatant;
2. 8mL of 0.5% potassium chloride hypotonic solution was added to the cell pellet, and the mixture was pipetted uniformly and placed in a 37 ℃ water bath for 30 min. Centrifuging at 1000rpm for 10min, and discarding supernatant;
3. slowly adding 5mL of stationary liquid (methanol: acetic acid = 3: 1) into the cell sediment, uniformly mixing, fixing in a water bath at 37 ℃ for 20min, centrifuging at 1000rpm for 10min, and discarding the supernatant;
4. adding 5mL of stationary liquid into the cell sediment, centrifuging at 1000rpm for 5min, removing the supernatant, leaving a little stationary liquid, and gently mixing the cells;
5. sucking cells by a liquid shifter, dripping the cells on a precooled glass slide, passing the cells on flame for several times, and naturally drying the cells; dyeing with Giemsa dye liquor for 20min, washing with running water, naturally drying, observing chromosome number with a microscope, selecting cover glass with good chromosome dispersion, multiple metaphase phases and regular morphology, sealing, and photographing to record results;
6. the chromosome number of the hybridoma cell is close to the sum of the chromosome numbers of two parent cells, namely the sum of the chromosome number (40 pieces) of the mouse spleen cell and the chromosome number (54-64) of the mouse myeloma cell NS-1;
the results of karyotyping of hybridoma cell line 7C11 obtained in example 1 are shown in FIG. 6, and the number of chromosomes was 94-103.
Claims (7)
1. A hybridoma cell strain secreting anti-human IgG3 monoclonal antibody is characterized in that:
the cell line was named: hybridoma cell line 7C 11; the culture is preserved in China center for type culture Collection with a preservation date of 2021, 11 months and 10 days, and the preservation number is CCTCC NO: C2021101.
2. the use of the hybridoma cell line secreting anti-human IgG3 monoclonal antibody of claim 1 in the preparation of human complement 7C11 detection reagents.
3. An anti-human IgG3 monoclonal antibody, characterized by:
the anti-human IgG1 monoclonal antibody is secreted and produced by the hybridoma cell line secreting anti-human IgG3 monoclonal antibody of claim 1.
4. A human IgG3 detection reagent, comprising:
comprising an anti-human IgG3 monoclonal antibody of claim 3.
5. A human IgG3 detection device, comprising:
comprising an anti-human IgG3 monoclonal antibody of claim 3.
6. A method for detecting human IgG3, comprising:
comprises the step of contacting a test sample with the anti-human IgG3 monoclonal antibody of claim 3 to determine whether the test sample contains human IgG 3.
7. The use of the anti-human IgG3 monoclonal antibody of claim 3 in the preparation of a human IgG3 detection reagent.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61124377A (en) * | 1984-11-22 | 1986-06-12 | Agency Of Ind Science & Technol | Novel hybridoma, preparation and use thereof |
| JPS62201596A (en) * | 1986-02-27 | 1987-09-05 | Shionogi & Co Ltd | Monoclonal antibody against human igg subclass, production thereof and cell system producing said monoclonal antibody |
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2022
- 2022-03-10 CN CN202210232046.0A patent/CN114874994A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61124377A (en) * | 1984-11-22 | 1986-06-12 | Agency Of Ind Science & Technol | Novel hybridoma, preparation and use thereof |
| JPS62201596A (en) * | 1986-02-27 | 1987-09-05 | Shionogi & Co Ltd | Monoclonal antibody against human igg subclass, production thereof and cell system producing said monoclonal antibody |
Non-Patent Citations (2)
| Title |
|---|
| HAJIGHASEMI F 等: "Generation and Characterization of Mouse Hybridomas Secreting Monoclonal Antibodies Specific for Human IgG3", 《AVICENNA J MED BIOTECHNOL》, vol. 1, no. 1, pages 21 * |
| 丁皓 等: "抗人IgG单克隆抗体制备和鉴定", 《上海医科大学学报》, vol. 21, no. 3, pages 165 - 169 * |
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