CN114805589B - Monoclonal antibody capable of simultaneously recognizing cow, goat and sheep antibodies - Google Patents

Monoclonal antibody capable of simultaneously recognizing cow, goat and sheep antibodies Download PDF

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CN114805589B
CN114805589B CN202210668484.1A CN202210668484A CN114805589B CN 114805589 B CN114805589 B CN 114805589B CN 202210668484 A CN202210668484 A CN 202210668484A CN 114805589 B CN114805589 B CN 114805589B
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sheep
monoclonal antibody
goat
antibodies
igg
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CN114805589A (en
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张春鸽
王格
赵巧辉
柴丹丹
徐延伟
李桂林
付光宇
杨增利
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Zhengzhou Immuno Biotech Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • C07K16/4208—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig
    • C07K16/4241—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig against anti-human or anti-animal Ig
    • 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
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56911—Bacteria
    • 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
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • 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
    • G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/35—Valency
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/08—RNA viruses
    • G01N2333/085—Picornaviridae, e.g. coxsackie virus, echovirus, enterovirus
    • G01N2333/09—Foot-and-mouth disease virus
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/23—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Brucella (G)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2469/00—Immunoassays for the detection of microorganisms
    • G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms

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Abstract

The invention relates to the technical field of in-vitro diagnosis, in particular to a monoclonal antibody capable of simultaneously identifying bovine, goat and sheep antibodies. The invention provides a monoclonal antibody, the heavy chain variable region of which has an amino acid sequence shown as SEQ ID No. 1. The monoclonal antibody of the invention can recognize bovine serum, goat serum, sheep serum and antibodies thereof; the antibody can also be used as a secondary antibody for development and detection of goat and sheep polyclonal antibodies and monoclonal antibodies; can also be used as the secondary antibody of the bovine and sheep immune antibody detection kit.

Description

Monoclonal antibody capable of simultaneously recognizing cow, goat and sheep antibodies
Technical Field
The invention relates to the technical field of in-vitro diagnosis, in particular to a monoclonal antibody capable of simultaneously identifying bovine, goat and sheep antibodies.
Background
Immunoglobulins refer to globulins having antibody activity or chemical structures similar to antibodies, primarily in blood and certain secretions. Immunoglobulins generally have antibody activity and bind specifically to the corresponding epitope, but are themselves an antigenic substance for other animals. The genetic background of immunoglobulin production varies, as does the antigenicity, and they can produce immune responses in xenogeneic, allogeneic and the same body, producing corresponding antibodies (anti-antibodies). Human immunoglobulins can be classified into five classes, igG, igA, igM, igD and IgE, depending on the antigen specificity of the heavy chain constant region of the immunoglobulin. Wherein IgG is the immunoglobulin with the highest content in serum and accounts for 75-80% of the total serum. The content of immunoglobulins in normal humans is relatively constant. Only in certain disease cases can the amount or quality of the immunoglobulins change.
The IgG species heterogeneity comes from the constant region (C region) of the immunoglobulin at 1/2 (about 105 amino acid residues) near the C-terminus of the L chain and at the 3/4 region or 4/5 region (about amino acids 119 to C-terminus) near the C-terminus of the H chain. The composition and arrangement of amino acids in this region are relatively constant in both Ig isotype L chains and H chains of the same class in animals of the same genus. Because of the presence of such species specificity, an immunoglobulin is caused to be heterogeneous among species, which causes an immune response among species.
When detecting IgG antibody concentration in vivo using ELISA methods, a secondary antibody is typically used.
The polyclonal antibody has the advantages of high coverage rate of human IgG protein sites, and basically no possibility of missed detection in the theoretical detection process. Meanwhile, the sensitivity of the self-production kit can be improved by utilizing the high affinity of the polyclonal antibody.
With the advantages of the application of polyclonal antibodies, development and research of polyclonal antibodies have been more in recent years, and mouse-goat monoclonal antibodies are often used as secondary antibodies in screening when the development of polyclonal antibodies is performed.
Sheep monoclonal antibodies have the advantage of strong affinity, and in recent years, development and research of sheep monoclonal antibodies are more, and when the sheep monoclonal antibodies are developed, mouse anti-sheep monoclonal antibodies are frequently used as secondary antibodies in screening.
Animal common disease vaccination, will affect antibody detection. The indirect detection kit can be used for secondary antibodies such as mouse-anti-sheep, mouse-anti-cow and the like. If there is a monoclonal antibody which can react with goat, sheep and cow at the same time, a kit can be developed, and the kit is universal for cow and sheep.
Disclosure of Invention
In view of this, the present invention provides monoclonal antibodies that can recognize bovine, caprine, ovine antibodies simultaneously, and the monoclonal antibodies of the present invention are suitable for the preparation of reagents and/or kits for cattle and sheep.
In order to achieve the above object, the present invention provides the following technical solutions:
the present invention provides monoclonal antibodies, heavy chain variable regions of which:
(I) Has an amino acid sequence shown as SEQ ID No. 1; and/or
Its light chain variable region:
(II) has an amino acid sequence shown as SEQ ID No. 2; and/or
Its heavy chain variable region and/or light chain variable region:
(III) an amino acid sequence having one or more residues substituted, deleted or added thereto as shown in (I) or (II), and having the same or similar function as the amino acid sequence shown in (I) or (II); and/or
(IV) an amino acid sequence having at least 80% homology with any of the amino acid sequences shown in (I) to (III);
the number of the said multiple is 2-30.
The invention also provides application of the monoclonal antibody in preparation of reagents and/or kits for identifying bovine, goat and/or sheep antibodies and/or serum.
In some embodiments of the invention, the bovine, caprine and/or ovine antibodies described above are derived from body fluids or secretions.
The invention also provides application of the monoclonal antibody in preparing a universal immunological detection reagent and/or a universal immunological detection kit for cattle and sheep.
The invention also provides application of the monoclonal antibody in preparation of a reagent and/or a kit for detecting cattle and sheep diseases.
In some embodiments of the invention, the bovine and ovine diseases in the above applications include foot-and-mouth disease and/or brucellosis.
In some embodiments of the invention, the above-described bovine and ovine diseases further include bacterial, viral and parasitic diseases of cattle and sheep; the bacterial diseases comprise clostridial diseases, pasteurellosis, salmonella diseases, listeriosis, tetanus and streptococcal diseases; the virus diseases comprise rinderpest, infectious bovine diarrhea, infectious bovine rhinotracheitis, bovine leukemia, bovine epidemic heat, bovine coronavirus disease, bluetongue disease, valvular heat, peste des petits ruminants disease, vaccinia and Niu Jiejie disease; the parasitic diseases comprise pyroworm disease, helminthiasis, bovine and sheep rhinomyiasis, taeniasis, ascariasis and enterobiasis.
The invention also provides a preparation method of the monoclonal antibody, which comprises the following steps:
step (a), immunizing a mouse by adopting bovine IgG as an immunogen;
step (b), taking spleen cells of the mice in the step (a) and fusing myeloma cells to obtain hybridoma cells;
step (c), screening and purifying the hybridoma cells in the step (b);
step (d) of obtaining the monoclonal antibody from the culture of the hybridoma cells of step (c);
the mice include BALB/c mice; and/or
The myeloma cells include myeloma cell NS1.
In some embodiments of the invention, the screening conditions in the above preparation method include: the monoclonal antibodies secreted by the hybridoma cells can specifically bind to bovine, caprine and/or ovine antibodies.
The invention also provides a reagent comprising the monoclonal antibody and acceptable auxiliary materials and/or auxiliary agents.
The invention also provides a kit comprising the monoclonal antibody or the reagent and acceptable auxiliary materials and/or auxiliary agents.
The invention also provides a device coated with the monoclonal antibodies or the reagents described above, and acceptable components.
The invention relates to a mouse monoclonal antibody for recognizing bovine serum, goat serum, sheep serum and antibodies thereof, which is characterized in that: the murine monoclonal antibody is designated as secondary antibody 4C-1, having a heavy chain variable region with the amino acid sequence shown in SEQ ID No.1 and a light chain variable region with the amino acid sequence shown in SEQ ID No. 2.
The invention discloses a mouse monoclonal antibody for recognizing bovine serum, goat serum, sheep serum and antibodies thereof, which is obtained by immunizing a BALB/c mouse by adopting bovine IgG as an immunogen, then fusing spleen and myeloma cells NS1, screening and purifying.
The invention has the following effects:
the monoclonal antibody of the invention has the advantage of universality, and the mouse monoclonal antibody can recognize bovine serum, goat serum, sheep serum and antibodies thereof. Can be used as a universal secondary antibody for development and detection of goat and sheep polyclonal antibodies and monoclonal antibodies. Can be used as a universal secondary antibody of a bovine and sheep immune antibody detection kit, including foot-and-mouth disease, brucellosis and the like.
1. The detection result of the titer of the mouse monoclonal antibody shows that the titer of the monoclonal antibody prepared by the invention is more than 1/256000 by adopting the enzyme-coated immune plates of bovine IgG, goat IgG and sheep IgG, and the monoclonal antibody shows higher titer;
2. ELISA detection results of the monoclonal antibody and the cow, goat and sheep antibodies show that the mouse anti-goat secondary antibody has weak reactivity to sheep antibody IgG and bovine serum IgG; the murine anti-sheep secondary antibody has weak reactivity to goat serum IgG and bovine serum IgG; the monoclonal antibody prepared by the invention has strong reactivity to sheep antibody IgG, goat serum IgG and bovine serum IgG.
Detailed Description
The invention discloses a monoclonal antibody capable of simultaneously recognizing cow, goat and sheep antibodies, and the technical parameters can be properly improved by the person skilled in the art by referring to the content of the invention. It is expressly noted that all such similar substitutions and modifications will be apparent to those skilled in the art, and are deemed to be included in the present invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those skilled in the relevant art that variations and modifications can be made in the methods and applications described herein, and in the practice and application of the techniques of this invention, without departing from the spirit or scope of the invention.
Unless otherwise indicated, all reagents used in the present invention are conventional and the apparatus and methods employed are conventional in the art. In the detection reagent and the detection kit provided by the invention, the raw materials and the reagents used can be purchased from the market.
The invention is further illustrated by the following examples:
example 1: preparation of murine monoclonal antibodies
1. Immunization of mice
Fully emulsifying bovine IgG with Freund's complete adjuvant, and immunizing female BALB/c mice with 5 weeks old in abdominal cavity, wherein the priming dose is 150 mug/mouse; the second and third immunizations were performed 21 days and 42 days after the first immunization, respectively, with an immunization dose of 70 μg/mouse. Blood is collected from the tail of the patient about 10 days after the third immunization, and serum titers are detected by a 96-well plate indirect method coated with bovine IgG.
2. Hybridoma cell preparation
Serum titers greater than 10 were detected by selective indirect method 4 The mice of (2) were subjected to intra-spleen booster immunization at a dose of 100. Mu.g/mouse. The spleen of the mice was fused with the myeloma cells NS1 of the mice at a ratio of 10:1 3 days after the booster immunization, and the fused cells were cultured on a DMEM medium (Gibco) containing HAT.
About 6-7 days after fusion, using 96-well plates coated with bovine IgG, goat IgG and sheep IgG, respectively adopting an indirect method to detect the specific antibody content in cell culture supernatant, selecting positive wells with the reaction OD value not lower than 0.5 in the three 96-well plates, and carrying out 3 rounds of subcloning by using a limiting dilution method, thereby finally respectively obtaining hybridoma cell strain 4C-1 capable of stably secreting anti-bovine IgG.
3. Murine monoclonal antibody purification
Injecting the obtained mouse hybridoma capable of stably secreting the monoclonal antibody into the abdominal cavity of a mouse, collecting ascites, and purifying by SPA to obtain the anti-mouse monoclonal antibody with the purity of more than 90%.
4. Titer detection of murine monoclonal antibodies
Bovine IgG, goat IgG, sheep IgG were diluted to 1 μg/mL with 0.05mmol/L, pH =9.6 CB buffer, 50 μl was added to each well of a 96 well enzyme-free plate (Corning) and coated overnight at 4 ℃. The next day was washed 3 times with PBST and then blocked with 1% Casein at 100. Mu.L/well for 2 hours at 37 ℃. Purified monoclonal antibody 4C-1 (concentrations of 5mg/mL each) was diluted in a ratio of 1:1000, 1:2000, 1:4000, 1:8000, 1:16000, 1:32000, 1:64000, … … with 0.05mmol/L, pH =9.6 CB buffer. The diluted antibodies were added to the enzyme-free plates coated with bovine IgG, goat IgG, sheep IgG, and the negative control wells were added with 0.05mmol/L, pH =9.6 CB buffer, 50 μl/well. After 30 minutes of reaction at 37℃the plates were washed 5 times with PBST, and after drying by pipetting, HRP-goat anti-mouse IgG (SIGMA) diluted 1:2000 was added, 100. Mu.L/well and reacted at 37℃for 30 minutes. The plate was washed 5 times with PBST, after drying, a normal enzyme-free substrate was added, 100. Mu.L/well was allowed to react at room temperature in the dark for 10 minutes, 50. Mu.L of sulfuric acid at a concentration of 0.1mol/L was added to terminate the reaction, and the absorbance at 450nm was measured. The test results are shown in Table 1 below:
TABLE 1 detection results
As can be seen from Table 1, the average OD value of the negative control is 0.01, and the bovine IgG, the goat IgG and the sheep IgG are coated by the enzyme-free plate for detection, and the titer of the monoclonal antibody reaches more than 1/256000, thus showing higher titer.
Example 2: ELISA detection results of monoclonal antibodies and bovine, goat and sheep antibodies
1. Magnetic particle coating
30 mu L of the uniformly mixed magnetic particle stock solution is taken and washed 5 times with 300 mu L of PBS buffer solution, then 10% glutaraldehyde is used for activating the magnetic particles for 1 hour, and then the activated magnetic particles are washed 2 times with the PBS buffer solution with pH of 7-8. The bovine, goat and sheep antibodies were all added to the beads in an amount of 0.3. Mu.g/human, and coated for 2 hours at 4 ℃. Finally, blocking with a blocking solution containing BSA was carried out for 2 hours.
2. HRP-labeled murine monoclonal antibody 4C-1
Monoclonal antibody 4C-1 was labeled with HRP in a mass ratio = 1.5:1 ratio. The HRP-labeled monoclonal antibody 4C-1 was diluted 1:4000 with diluent.
3. The magnetic particle chemiluminescence detection is carried out by a direct method. The test results are shown in Table 2 below:
TABLE 2 detection results
Analysis of results:
1. the detection result of the mouse anti-goat secondary antibody shows that the mouse anti-goat secondary antibody reacts with goat IgG, the detection luminescence value is more than 100000, the detection luminescence value is lower than 5000, and the mouse anti-goat secondary antibody only reacts with goat IgG and can be regarded as non-reaction with sheep IgG and cow IgG;
2. the detection result of the mouse anti-sheep secondary antibody shows that the mouse anti-sheep secondary antibody reacts with sheep IgG, the detection luminescence value is more than 100000, the detection luminescence value is more than goat IgG and cow IgG, and the detection luminescence value is lower than 5000, which indicates that the mouse anti-sheep secondary antibody only reacts with sheep IgG and can be regarded as non-reaction with sheep IgG and cow IgG;
3. the secondary antibody 4C-1 screened by the invention reacts with goat IgG, sheep IgG and cow IgG, and the detection luminescence values are more than 100000, which indicates that the secondary antibody 4C-1 reacts with goat, sheep and cow IgG.
The murine monoclonal antibody obtained by the invention is named as secondary antibody 4C-1, and the amino acid sequences of the heavy chain variable region and the light chain variable region are as follows:
amino acid sequence of heavy chain variable region (SEQ ID No. 1):
QVQLKQSGPELVKPGASVKMSCKASGYTFTSYVIHWVKQKPGQGLEWIGYINPYNDNTEYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCARREGGYYVFTYWGQGTLVTVSA
amino acid sequence of light chain variable region (SEQ ID No. 2):
EMVLTQSPALMAASPGEKVTITCSVSSSISSSNFHWYQQKSETSPKPWIYGTSNLASGVPVRFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSYPLTFGGGTKLEIK
the foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Sequence listing
<110> Imeno Biotechnology Co., zhengzhou
<120> monoclonal antibody capable of recognizing bovine, goat and sheep antibodies simultaneously
<130> MP21028305
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 119
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 1
Gln Val Gln Leu Lys Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Val Ile His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Tyr Ile Asn Pro Tyr Asn Asp Asn Thr Glu Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ser Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Glu Gly Gly Tyr Tyr Val Phe Thr Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ala
115
<210> 2
<211> 108
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 2
Glu Met Val Leu Thr Gln Ser Pro Ala Leu Met Ala Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Ile Thr Cys Ser Val Ser Ser Ser Ile Ser Ser Ser
20 25 30
Asn Phe His Trp Tyr Gln Gln Lys Ser Glu Thr Ser Pro Lys Pro Trp
35 40 45
Ile Tyr Gly Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu
65 70 75 80
Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Tyr Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105

Claims (4)

1. The monoclonal antibody is characterized in that the heavy chain variable region of the monoclonal antibody has an amino acid sequence shown as SEQ ID No. 1; and
the light chain variable region has an amino acid sequence shown as SEQ ID No. 2.
2. Use of a monoclonal antibody according to claim 1 for the preparation of an immunological detection reagent and/or kit for simultaneous identification of cattle, goats, sheep.
3. A universal reagent for simultaneous identification of cattle, goats and sheep comprising a monoclonal antibody according to claim 1 and acceptable adjuvants and/or adjuvants.
4. A universal kit for simultaneous identification of cattle, goats, sheep comprising a universal reagent according to claim 3.
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RU2377299C1 (en) * 2008-06-10 2009-12-27 Государственное научное учреждение Всероссийский научно-исследовательский институт экспериментальной ветеринарии им. Я.Р. Коваленко РАСХН (ВИЭВ) STRAIN 5A10 OF PERMANENT HYBRIDOMAL LINE OF CELLS OF MOUSE Mus. musculus - PRODUCENT OF MONOCLONAL ANTIBODIES TO IgG OF CATTLE
CN104099300A (en) * 2013-11-22 2014-10-15 沈阳农业大学 Hybridoma cell line able to secrete anti-bovine immunoglobulin IgG monoclonal antibody and application thereof
CN109507421A (en) * 2018-12-07 2019-03-22 广西大学 Water resistant ox IgG monoclonal antibody cell strain and its preparation method and application
CN111896750A (en) * 2020-08-07 2020-11-06 杭州都林生物科技有限公司 Bovine immunoglobulin (IgG) double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method
CN112010981A (en) * 2020-09-10 2020-12-01 南京妙迪生物科技有限公司 Mouse anti-human IgG monoclonal antibody
CN113004413A (en) * 2019-12-19 2021-06-22 中国农业大学 Monoclonal antibody of porcine IgG3, epitope peptide specifically recognized by monoclonal antibody and application of epitope peptide

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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