CN117050168B - Monoclonal antibody against helicobacter pylori UreA protein and application thereof - Google Patents
Monoclonal antibody against helicobacter pylori UreA protein and application thereof Download PDFInfo
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- C07K16/12—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria
- C07K16/1203—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria
- C07K16/121—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria from Helicobacter (Campylobacter) (G)
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- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
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- 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
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- 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
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
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- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
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Abstract
The invention discloses a monoclonal antibody of anti-helicobacter pylori UreA protein and application thereof, in particular relates to a monoclonal antibody of anti-UreA protein or antigen binding fragment thereof, wherein the monoclonal antibody or antigen binding fragment thereof comprises a heavy chain variable region CDR1 shown in SEQ ID NO. 1, a heavy chain variable region CDR2 shown in SEQ ID NO. 2, a heavy chain variable region CDR3 shown in SEQ ID NO. 3 and SEQ ID NO:9, the light chain variable region CDR1, SEQ ID NO:10, the light chain variable region CDR2, SEQ ID NO:11, and a light chain variable region CDR3. The invention also provides nucleotide molecules, vectors, host cells, methods and uses encoding the monoclonal antibodies or antigen binding fragments thereof.
Description
Technical Field
The invention belongs to the technical field of biology, and relates to a monoclonal antibody for resisting helicobacter pylori UreA protein and application thereof.
Background
The helicobacter is a fungus of the class epsilon-Proteus, order Campylobacter, family Campylobacter. Also known as helicobacter pylori. Helicobacter pylori HP (Helicobocton Pyloni), which was first discovered by Swedish scholars from human gastric mucosa specimens, is also a subject of recent domestic research. The world health organization first included helicobacter pylori in a list of carcinogens on day 10 and 27 of 2017.
Helicobacter pylori is a gram-negative helicobacter found on the gastric mucosa and grows in a micro-aerobic environment, positive for oxidase and catalase, has smooth cell walls and 1-5 flagella, and the latter is sleeved in a sheath and the tail end is spherical. Helicobacter infection is mainly helicobacter pylori (Helicobacter pylori, hp) infection. Helicobacter pylori is gram-negative, and the thallus is arc, S-shaped or spiral, has flagella, and is active in movement and free of spores. The bacterium has weak resistance to the external environment and is sensitive to both drying and heat. Various commonly used disinfectants are easy to kill. The pathogenicity of helicobacter pylori is related to factors such as toxins produced by it, enzymes with toxic effects destroying gastric mucosa and causing the body to produce inflammation and immune response. Helicobacter pylori infection can trigger inflammation and immune response, cell degeneration, necrosis and inflammatory cell infiltration are seen in the gastric mucosa it infects, and specific antibodies are detected in serum. Helicobacter pylori is associated with gastritis, peptic ulcer, gastric cancer, gastric mucosa-associated lymphoid tissue lymphoma (MALT lymphoma), non-steroidal anti-inflammatory drug-associated gastropathy functional dyspepsia, gastroesophageal reflux disease, and other diseases.
Disclosure of Invention
In order to solve the technical problems existing in the real world, the invention aims to provide a monoclonal antibody for resisting helicobacter pylori UreA protein.
In a first aspect, the invention provides a monoclonal antibody against the helicobacter pylori UreA protein, the monoclonal antibody comprising monoclonal antibody 7G8.
The monoclonal antibody 7G8 comprises: SEQ ID NO:1, a heavy chain variable region CDR1, SEQ ID NO:2, a heavy chain variable region CDR2, SEQ ID NO:3, a heavy chain variable region CDR3, SEQ ID NO:9, the light chain variable region CDR1, SEQ ID NO:10, the light chain variable region CDR2, SEQ ID NO:11, and a light chain variable region CDR3.
In certain embodiments, the antibodies and functional fragments thereof provided herein comprise one or more amino acid residue substitutions in one or more of the CDR sequences. In certain embodiments, the affinity variants comprise no more than 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 substitutions in total in the CDR sequence.
Further, the monoclonal antibody 7G8 further comprises a polypeptide having a sequence identical to SEQ ID NO: 4. 5, 6, 7, and a heavy chain variable region framework region FR1, FR2, FR3, FR4 having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 12. 13, 14, 15, and at least 90% sequence identity to the light chain variable region framework regions FR1, FR2, FR3, FR4.
In certain embodiments, the antibodies and functional fragments thereof provided herein comprise one or more amino acid residue substitutions in one or more of the FR sequences. In certain embodiments, the affinity variants comprise no more than 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 substitutions in total in the FR sequence.
Further, the monoclonal antibody 7G8 heavy chain variable region has a sequence identical to SEQ ID NO:8, preferably a heavy chain variable region having at least 90% sequence identity, preferably 95% sequence identity, to the amino acid sequence set forth in SEQ ID NO:16, preferably a light chain variable region having at least 90% sequence identity, preferably 95% sequence identity.
In a specific embodiment, the heavy chain variable region of monoclonal antibody 7G8 has the amino acid sequence shown in SEQ ID NO. 8; the light chain variable region of the monoclonal antibody 7G8 has an amino acid sequence shown in SEQ ID NO. 16.
Further, the monoclonal antibody further comprises a heavy chain constant region and a light chain constant region.
The term "antibody" includes reference to immunoglobulin molecules and antigen binding portions of immunoglobulin (Ig) molecules, i.e., molecules that contain antigen binding sites that specifically bind to (immunoreact with) an antigen. "SpecialBy "specifically bind" or "immunoreact with" … or "immunospecifically bind" is meant that the antibody reacts with one or more epitopes of the desired antigen and does not react with other polypeptides or bind with much lower affinity (Kd>10 -6 ). Antibodies include, but are not limited to, polyclonal antibodies, monoclonal antibodies, chimeric antibodies, fully human antibodies, domain antibodies, single chains, fab and F (ab') 2 fragments, scFv and Fab expression libraries.
Further, the monoclonal antibody may be an afucosylated antibody, or a partially afucosylated antibody.
In certain specific embodiments, the monoclonal antibodies further comprise an immunoglobulin constant region, optionally a heavy chain and/or a light chain constant region. In certain specific embodiments, the heavy chain constant region comprises a CH1, hinge, and/or CH2-CH3 region (or optionally a CH2-CH3-CH4 region). The immunoglobulin constant region of the monoclonal antibody provided by the invention is consistent with, differs from the wild-type immunoglobulin constant region by one mutation or differs from the wild-type immunoglobulin constant region by a plurality of mutations.
In a second aspect the invention provides a nucleic acid molecule encoding a monoclonal antibody or a functional fragment thereof as hereinbefore described.
Further, the functional fragment of the monoclonal antibody includes a nucleic acid functional fragment or an amino acid functional fragment.
Further, the nucleic acid functional fragments also include corresponding nucleic acid fragments of amino acid fragments that the antibody is capable of functioning, including RNA and DNA fragments, including, but not limited to mRNA, tRNA, rRNA, snRNA, hRNA, antisense RNA, tCRNA, dsRNA, SCRNA, catalytically active RNA, various viral RNAs, single-stranded DNA, closed-loop DNA, ligated DNA, and the like.
In a third aspect the present invention provides a recombinant expression vector comprising a nucleic acid molecule as hereinbefore described.
Further, the recombinant expression vector includes a lentiviral vector, an adenovirus vector, an adeno-associated virus (AAV) vector, a retrovirus vector, a plasmid, a DNA vector, an mRNA vector, a transposon-based vector, or an artificial chromosome.
Further, the plasmid includes a eukaryotic cell expression vector.
The term "vector" refers to a nucleic acid molecule that may be engineered to contain a polynucleotide or polynucleotides that may be amplified in a host cell for cloning. Vectors include, but are not limited to: a single-stranded, double-stranded or partially double-stranded nucleic acid molecule; nucleic acid molecules comprising one or more free ends, without free ends (e.g., circular); a nucleic acid molecule comprising DNA, RNA, or both; and other polynucleotide species known in the art. One type of vector is a "plasmid," which refers to a circular double-stranded DNA loop into which additional DNA fragments may be inserted. Certain vectors are capable of autonomous replication in the host cell into which they are introduced. Other vectors integrate into the host cell genome upon introduction into the host cell and thereby replicate together with the host genome.
In a fourth aspect the invention provides an engineered host cell comprising a monoclonal antibody as described hereinbefore or a functional fragment thereof, or a nucleic acid molecule as described hereinbefore, a recombinant expression vector as described hereinbefore.
Further, the engineered host cell includes a prokaryotic cell or a eukaryotic cell.
Further, the prokaryotic cells include bacteria, actinomycetes, cyanobacteria, mycoplasma, chlamydia, rickettsia.
Further, the bacteria include gram-positive bacteria and gram-negative bacteria.
Further, the eukaryotic cells include mammalian cells, insect cells, plant cells other than cyanobacteria, fungal cells, yeast cells.
Further, the mammalian cells include fibroblasts, lymphocytes, epithelial cells, and myeloblasts.
Further, the host cell includes a hybridoma cell.
The term "host cell" refers to a prokaryotic cell, such as a bacterial cell; or lower eukaryotic cells, such as yeast cells; or higher eukaryotic cells, such as mammalian cells. Representative examples are: coli, streptomyces; bacterial cells such as salmonella typhimurium; fungal cells such as yeast; a plant cell; insect cells such as Drosophila S2 or Sf9; animal cells such as CHO, COS or Bowes melanoma cells, etc.
In a fifth aspect the invention provides an antibody conjugate comprising a complex of a monoclonal antibody as hereinbefore described or a functional fragment thereof, directly or indirectly conjugated to a detectable label.
As used herein, the term "detectable label" refers to any moiety that generates a detectable signal through a change in an optical, electrical, or other physical indicator of the state of a molecule coupled to the moiety. Such physical indicators include spectroscopic, photochemical, biochemical, immunochemical, electromagnetic, radiochemical and chemical means such as, but not limited to, fluorescence, chemiluminescence and the like. When used in reference to a labeled detection agent, a "direct label" is a detectable label attached to the detection agent by any means. An "indirect label" is a detectable label that specifically binds to a detection agent when used in reference to a labeled detection agent. Thus, the indirect label comprises the following moieties: which is a specific binding partner for the detector moiety. Biotin and avidin are examples of such moieties employed, for example, by contacting a biotinylated antibody with a labeled avidin to produce an indirectly labeled antibody.
Further, the functional fragment of the monoclonal antibody also includes a nucleic acid-like functional fragment that can bind to a nucleic acid molecule encoding the antigen helicobacter pylori.
In a sixth aspect, the invention provides a product for the detection of helicobacter pylori, said product comprising a monoclonal antibody as defined above or a functional fragment thereof, a nucleic acid molecule as defined above, a recombinant expression vector as defined above, an engineered host cell as defined above or an antibody conjugate as defined above.
Further, the product includes a kit, a test paper, a nucleic acid membrane strip, or a chip.
In a seventh aspect, the invention provides a composition comprising a monoclonal antibody as described above or a functional fragment thereof, a nucleic acid molecule as described above, a recombinant expression vector as described above, an engineered host cell as described above.
According to an eighth aspect of the present invention there is provided a method of producing a monoclonal antibody as hereinbefore described, the method comprising the steps of:
a) Providing a nucleic acid molecule as described above or a recombinant expression vector as described above;
b) Transforming a host cell with the substance of step a);
c) Culturing the host cell obtained in step b) under suitable conditions;
d) The monoclonal antibody is obtained by separation and purification in a host cell culture solution.
In a ninth aspect, the present invention provides a method for detecting helicobacter pylori protein or a nucleic acid molecule encoding the same in a sample to be tested, at a non-diagnostic and non-therapeutic destination, the method comprising the steps of: contacting a test sample with the monoclonal antibody or functional fragment thereof described above, or contacting a test sample with the antibody conjugate described above; detecting the formation of complexes of helicobacter pylori proteins or nucleic acid molecules encoding the same with the monoclonal antibodies described above, or the antibody conjugates described above.
In a tenth aspect the present invention provides a method of preparing an engineered host cell as hereinbefore described, said method comprising the steps of: the monoclonal antibodies or functional fragments thereof, the nucleic acid molecules, or the recombinant expression vectors are introduced into host cells.
In an eleventh aspect, the present invention provides a method for specifically inhibiting helicobacter pylori, the method comprising the steps of: the monoclonal antibody or a functional fragment thereof, or the nucleic acid molecule, or the recombinant expression vector, is introduced into a cell of an organism to inhibit the activity of helicobacter pylori protein by expressing the monoclonal antibody or the functional fragment thereof.
The term "helicobacter pylori" refers to helicobacter pylori, a gram-negative helicobacter found on the gastric mucosa, growing in a micro-aerobic environment, being positive for oxidase and catalase, having smooth cell walls and 1-5 flagella, the latter being sheathed in a sheath and having a globular end. The helicobacter is a gram negative bacterium, is spiral, and has a thin and long body of 5-50 μm. They are extremely active, moving in a spiral. The helicobacter bacteria can be classified into aerobic, absolute anaerobic and anaerobic bacteria. They typically multiply when the wastewater or wastewater sample is in a transition between aerobic and anaerobic. The fungus has high diversity. While some spirochetes are pathogenic, such as pathogens that cause syphilis, most are autogenous in the soil. They are introduced into the activated sludge process by means of influent and permeate
In a twelfth aspect, the invention provides the use of a monoclonal antibody as defined above or a functional fragment thereof, a nucleic acid molecule as defined above, a recombinant expression vector as defined above, an engineered host cell as defined above, or an antibody conjugate as defined above for the detection of the helicobacter pylori UreA protein or fragment thereof.
Further, the helicobacter pylori UreA protein or a fragment thereof includes a UreA protein, a UreA protein fragment, a nucleic acid molecule encoding a UreA protein fragment.
In a thirteenth aspect, the invention provides the use of a monoclonal antibody as defined above or a functional fragment thereof, a nucleic acid molecule as defined above, a recombinant expression vector as defined above, a modified host cell as defined above for the preparation of a product for diagnosing helicobacter pylori infection.
In a fourteenth aspect, the invention provides the use of a monoclonal antibody as defined above or a functional fragment thereof, a nucleic acid molecule as defined above, a recombinant expression vector as defined above, a modified host cell as defined above for the manufacture of a medicament for the prophylaxis or treatment of helicobacter pylori infection.
In a fifteenth aspect, the invention provides the use of a monoclonal antibody as defined above or a functional fragment thereof, a nucleic acid molecule as defined above, a recombinant expression vector as defined above, a host cell as defined above, for the manufacture of a medicament for modulating the activity or level of a helicobacter pylori protein.
Drawings
FIG. 1 is a diagram showing the result of electrophoresis for detecting monoclonal antibody 7G8;
FIG. 2 is a graph of HPLC results for detecting monoclonal antibody 7G8;
FIG. 3 is a graph showing the results of ELISA detection of the binding activity of monoclonal antibody 7G8.
Detailed Description
Before describing the present methods, it is to be understood that this invention is not limited to the particular methods and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.
EXAMPLE 1 screening of monoclonal antibodies
1. Recombinant expression of immunogens
Synthesizing a helicobacter pylori UreA protein sequence, and constructing a pEM5.1 vector; extracting the plasmid for transfection; transfecting HEK293 cells, and culturing the cells for 7 days; and (3) harvesting supernatant, purifying by a Ni column, concentrating a replacement buffer solution to obtain recombinant helicobacter pylori UreA protein, and obtaining a recombinant helicobacter pylori UreA protein sequence source Uniprot. The sequence is shown in SEQ ID NO. 17, and is specifically shown in Table 1.
TABLE 1 recombinant helicobacter pylori UreA protein sequence
2. Immunization
A second immunization with complete Freund's adjuvant, 100 μg each, with a total dose of 0.5 ml/dose per abdominal cavity, 3 weeks apart; the second time was boosted with Freund's incomplete adjuvant at a dose of 50. Mu.g/0.5 ml/dose, with a third immunization at 2 week intervals; cell fusion was prepared 10 days after the third injection;
feeder cells were taken at a rate of 10 5 Well use, plating 10 the day prior to fusion 5 Mu.l/well; taking mouse immune spleen cells and prepared myeloma cells to fuse with a fusion agent PEG, paving the mixture into a 96 cell culture plate added with feeder cells,100 μl/well.
3. Selection and cloning of hybridoma cells
Screening positive holes by ELISA detection method, and paving recombinant helicobacter pylori UreA protein overnight; washing the plate, adding skimmed milk powder, sealing, and standing at 37deg.C for 1 hr; washing the plate, adding 100 μl of 96-well culture supernatant, and incubating at 37deg.C for 1 hr; washing the plate, adding HRP-labeled goat anti-mouse secondary antibody, and incubating for 30min at 37 ℃; washing the plate, adding a color development liquid, developing for 10min, adding a stop solution, and reading the value of OD 450; and screening the cell strain with high expression quantity for subcloning.
4. Sequence fishing
Cells were collected, total RNA was extracted using Trizol, and cDNA was generated by reverse transcription using oligo (dT) 20 as a primer. And then amplifying the heavy chain variable region genes by using specific primer PCR. After the PCR product is purified by electrophoresis, the PCR product is inserted into a vector through TA cloning, and is transformed, and positive clones are selected for sequencing.
5. Experimental results
Screening out monoclonal antibody 7G8 of helicobacter pylori UreA protein targeting helicobacter pylori UreA protein, the sequence is shown in Table 2.
TABLE 2 amino acid sequence of monoclonal antibody 7G8
EXAMPLE 2 functional Studies of monoclonal antibody 7G8
1. Expression and purification of monoclonal antibodies
(1) The screened sequences were chemically synthesized and cloned into eukaryotic expression vectors.
(2) The plasmid was amplified and extracted.
(3) The plasmid encoding the antibody was transiently transfected into mammalian cells HEK293.
(4) Collecting the supernatant, and purifying by using an affinity chromatography method to obtain the monoclonal antibody.
(5) The result showed that the amount of the purified monoclonal expression was 232mg/L.
2. Physicochemical property detection of monoclonal antibody
(1) Gel electrophoresis detection
A. Sample preparation
Mixing 20 μl sample with 5 μl 5×reduction buffer, heating at 95deg.C for 5min, and cooling; mu.L of the sample was mixed with 5. Mu.L of 5 Xnon-reducing buffer.
B. Electrophoresis
Preparing gel, adding proper amount of electrophoresis buffer solution, adding sample, and performing electrophoresis.
C. Dyeing and decoloring
After electrophoresis, taking gel, putting the gel into a proper amount of coomassie brilliant blue staining solution, and staining the gel for 1 hour or more at room temperature; pouring out the dyeing liquid, adding a proper amount of coomassie brilliant blue dyeing and decolorizing liquid at room temperature for 4-24 hours. After the completion of the decoloring, ddH was used 2 O soaking, referring to Marker protein, comparing with undyed gel, cutting gel of desired protein component, and collecting. The protein to be purified is then separated from the gel.
3. Experimental results
The results are shown in FIG. 1, wherein the left-right stripes are marker and restoration stripes respectively; the electrophoresis result graph shows that the monoclonal antibody 7G8 is expressed successfully.
(2) HPLC detection of purity of monoclonal antibodies
A. Flow matching
Dipotassium phosphate, tripotassium phosphate and potassium chloride are added into about 900mL of purified water, stirred and dissolved, the volume is fixed to 1L, and the pH value is determined to be 6.2+/-0.1 by measuring with a pH meter. Filtering with 0.22 μm filter membrane, and storing at room temperature.
B. Sample preparation
System applicability samples: MIL62 standard is diluted to 2mg/mL with mobile phase;
test article: the sample to be tested was diluted to 2mg/mL with mobile phase.
The detection was performed under conventional chromatographic conditions.
C. Experimental results
As shown in FIG. 2, the results of the liquid phase detection showed that the detection purity of the monoclonal antibody 7G8 was greater than 95%.
(3) Detection of binding Activity of monoclonal antibodies
A. Coating: the antigen UreA protein was diluted to 2. Mu.g/ml with coating solution, mixed well, added to 96-well coating plate, 100. Mu.l/well, blocked with a blocking film, and overnight at 4 ℃.
B. The plate washer is washed 3 times, no liquid can remain on the plate for the last time, and the liquid on the surface of the plate is beaten by the absorbent paper.
C. Closing: 5% milk powder (0.5 g milk powder in 10mL DPBS), 300. Mu.L/well, and incubated at 37℃for 1h, and plates were washed 3 times according to step 2).
D. The antibodies were subjected to gradient dilution, 100. Mu.L/well, reaction at 37℃for 1h, and plate washing was performed 3 times according to step B.
E. Adding a secondary antibody: the plates were washed 3 times according to step B with DPBS diluted 1:2000, added to 96-well plates, 100. Mu.L/well, reacted at 37℃for 1 h.
F. Color development: TMB was added thereto, 100. Mu.L/well, and the mixture was developed at room temperature in a dark place for 10 minutes.
G. And (3) terminating: add 2N H 2 SO 4 100. Mu.L/well.
H. The detection of the OD450 is carried out by an enzyme-labeled instrument within 10min.
I. Experimental results
The results are shown in FIG. 3, which shows that monoclonal antibody 7G8 was able to bind specifically to the UreA protein and exhibited a concentration dependence with an EC50 of 0.09152. Mu.g/mL.
The above description of the embodiments is only for the understanding of the method of the present invention and its core ideas. It should be noted that it will be apparent to those skilled in the art that several improvements and modifications can be made to the present invention without departing from the principle of the invention, and these improvements and modifications will fall within the scope of the claims of the invention.
Claims (28)
1. A monoclonal antibody against the helicobacter pylori UreA protein, characterized in that the monoclonal antibody comprises monoclonal antibody 7G8;
the monoclonal antibody 7G8 comprises: SEQ ID NO:1, a heavy chain variable region CDR1, SEQ ID NO:2, a heavy chain variable region CDR2, SEQ ID NO:3, a heavy chain variable region CDR3, SEQ ID NO:9, the light chain variable region CDR1, SEQ ID NO:10, and the light chain variable region CDR2 shown in SEQ ID NO:11, and a light chain variable region CDR3.
2. The monoclonal antibody of claim 1, wherein the monoclonal antibody 7G8 further comprises a polypeptide having a sequence identical to SEQ ID NO: 4. 5, 6, 7, and a heavy chain variable region framework region FR1, FR2, FR3, FR4 having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 12. 13, 14, 15, and at least 90% sequence identity to the light chain variable region framework regions FR1, FR2, FR3, FR4.
3. The monoclonal antibody of claim 1 or 2, wherein the monoclonal antibody 7G8 heavy chain variable region has a sequence identical to SEQ ID NO:8, and at least 90% sequence identity to the amino acid sequence set forth in seq id no.
4. A monoclonal antibody according to claim 3, wherein the monoclonal antibody 7G8 heavy chain variable region has a sequence identical to SEQ ID NO:8, and at least 95% sequence identity to the amino acid sequence set forth in seq id no.
5. The monoclonal antibody of claim 1 or 2, wherein the light chain variable region has a sequence identical to SEQ ID NO:16, and at least 90% sequence identity to the amino acid sequence set forth in seq id no.
6. The monoclonal antibody of claim 5, wherein the light chain variable region has a sequence identical to SEQ ID NO:16, and at least 95% sequence identity to the amino acid sequence set forth in seq id no.
7. The monoclonal antibody of claim 1 or 2, further comprising a heavy chain constant region and a light chain constant region.
8. The monoclonal antibody of claim 7, wherein the monoclonal antibody is an afucosylated antibody or a partially afucosylated antibody.
9. A nucleic acid molecule encoding the monoclonal antibody of any one of claims 1-8.
10. A recombinant expression vector comprising the nucleic acid molecule of claim 9.
11. The recombinant expression vector of claim 10, wherein the recombinant expression vector comprises an adenovirus vector, an adeno-associated virus (AAV) vector, a retrovirus vector, a DNA vector, an mRNA vector.
12. The recombinant expression vector of claim 11, wherein the DNA vector comprises a plasmid, a transposon-based vector, or an artificial chromosome, and wherein the plasmid comprises a eukaryotic cell expression vector.
13. An engineered host cell comprising the monoclonal antibody of any one of claims 1-8, or the nucleic acid molecule of claim 9, the recombinant expression vector of any one of claims 10-12.
14. The engineered host cell of claim 13, wherein the engineered host cell comprises a prokaryotic cell or a eukaryotic cell.
15. The engineered host cell of claim 14, wherein the prokaryotic cell comprises bacteria, mycoplasma, chlamydia, rickettsia.
16. The engineered host cell of claim 15, wherein the bacteria comprise gram positive bacteria, gram negative bacteria.
17. The engineered host cell of claim 14, wherein the eukaryotic cell comprises a mammalian cell, an insect cell, a plant cell other than cyanobacteria, a fungal cell.
18. The engineered host cell of claim 17, wherein the mammalian cell comprises a fibroblast, a lymphocyte, an epithelial cell, a myeloblast.
19. The engineered host cell of claim 13, wherein the host cell comprises a hybridoma cell.
20. An antibody conjugate comprising a complex of the monoclonal antibody of any one of claims 1-8 directly or indirectly conjugated to a detectable label.
21. A product for detecting helicobacter pylori, the product comprising the monoclonal antibody of any one of claims 1-8, the nucleic acid molecule of claim 9, the recombinant expression vector of any one of claims 10-12, the engineered host cell of any one of claims 13-19, or the antibody conjugate of claim 20.
22. The product of claim 21, comprising a kit, a test strip, a nucleic acid membrane strip, or a chip.
23. A composition comprising the monoclonal antibody of any one of claims 1-8, the nucleic acid molecule of claim 9, the recombinant expression vector of any one of claims 10-12, the engineered host cell of any one of claims 13-19.
24. A method of producing a monoclonal antibody according to any one of claims 1-8, comprising the steps of:
a) Providing the nucleic acid molecule of claim 9 or the recombinant expression vector of any one of claims 10-12;
b) Transforming a host cell with the substance of step a);
c) Culturing the host cell obtained in step b) under suitable conditions;
d) Isolation and purification in a host cell culture medium to obtain the monoclonal antibody of any one of claims 1-8.
25. A method of making the engineered host cell of any one of claims 13-19, comprising the steps of: introducing the monoclonal antibody of any one of claims 1-8, the nucleic acid molecule of claim 9, or the recombinant expression vector of any one of claims 10-12 into a host cell.
26. Use of the monoclonal antibody of any one of claims 1-8, the nucleic acid molecule of claim 9, the recombinant expression vector of any one of claims 10-12, the engineered host cell of any one of claims 13-19 for the preparation of a product for diagnosing helicobacter pylori infection.
27. Use of the monoclonal antibody of any one of claims 1-8, the nucleic acid molecule of claim 9, the recombinant expression vector of any one of claims 10-12, the engineered host cell of any one of claims 13-19 in the preparation of a medicament for preventing or treating helicobacter pylori infection.
28. Use of the monoclonal antibody of any one of claims 1-8, the nucleic acid molecule of claim 9, the recombinant expression vector of any one of claims 10-12, the engineered host cell of any one of claims 13-19 in the preparation of a medicament for modulating activity or level of helicobacter pylori protein.
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