Fully human anti-Der p1 full-molecule IgG antibody and application thereof

Abstract

The invention provides a full-human anti-Der p1 full-molecule IgG antibody and application thereof, and relates to the field of biological pharmacy. The full-human anti-Der p1 full-molecule IgG antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequences of the antigen complementary determining regions CDR of the light chain variable region are respectively as shown in SEQ ID NO.15, SEQ ID NO.16 and SEQ ID NO.17; the amino acid sequences of the antigen complementary determining region CDR of the heavy chain variable region are respectively shown as SEQ ID NO. 18, SEQ ID NO. 19 and SEQ ID NO. 20. The full-human anti-Der p1 full-molecule IgG antibody provides high specificity, high affinity, low immunogenicity and potential immunomodulatory function through the specific CDR design, and can provide a new treatment and diagnosis means for allergic diseases caused by dust mites.

Description

Technical Field

[0001] The present invention relates to the field of biopharmaceuticals, and in particular to a fully humanized anti-Der p 1 whole-molecule IgG antibody and an application thereof. Background Art

[0002] Allergic diseases are an increasingly serious health problem worldwide, and dust mites, as one of the most important allergens, pose a huge challenge to public health. Allergic diseases caused by dust mites include asthma, allergic rhinocorticitis, atopic dermatitis, etc., which seriously affect the quality of life of patients.

[0003] Among the existing technologies, research on dust mite allergens has made some progress. 115 types of mite allergens can be retrieved on the IUIS allergen naming website, among which Dermatophagoides pteronyssinus 1 (Der p 1) is the most important HDM allergen. It not only induces IgE reactions, but also has cysteine ​​protease properties, which can affect the occurrence of allergic asthma. Der p 1 triggers a proteolytic activation cascade in the dust mite digestive tract, leading to the maturation of multiple allergens, which trigger allergic reactions when dust mite fecal particles reach the airway mucosa.

[0004] However, the existing technology has some defects in the research and application of dust mite allergens. First, most of the antibodies against Der p 1 are mainly used for diagnosis and scientific research, and most of them are mouse antibodies or IgE antibodies. These antibodies may have immunogenicity and cross-reactivity problems in clinical applications. Secondly, the extraction and purification methods of Der p 1 have not been fully optimized, which affects the development of recombinant allergens, hypoallergenic derivatives and preventive vaccines. In addition, the effect of Der p 1 on airway epithelial cells and its specific mechanism of action in allergic reactions still need to be further clarified.

[0005] In summary, the existing technologies have the following major defects in the research and application of dust mite allergens: first, there is a lack of highly efficient and specific fully human antibodies; second, the extraction and purification technology of dust mite allergens needs to be further improved; and third, there is insufficient understanding of the mechanism of action of dust mite allergens in allergic reactions. These defects limit the application of dust mite allergens in clinical diagnosis and treatment, and also affect the development of new vaccines and drugs.

[0006] In view of this, the present invention is proposed. Summary of the invention

[0007] The purpose of the present invention is to provide a fully human anti-Der p 1 whole molecule IgG antibody and its application. The fully human anti-Der p 1 whole molecule IgG antibody has the advantages of high specificity, high affinity, low immunogenicity and potential immunoregulatory function.

[0008] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:

[0009] In a first aspect, the present invention provides a fully human anti-Der p 1 whole molecule IgG antibody, wherein the fully human anti-Der p 1 whole molecule IgG antibody comprises a light chain variable region and a heavy chain variable region;

[0010] The amino acid sequences of the antigen complementary determining regions (CDRs) of the light chain variable regions are shown in SEQ ID NO.15, SEQ ID NO.16 and SEQ ID NO.17 respectively;

[0011] The amino acid sequences of the antigen complementary determining regions (CDRs) of the heavy chain variable regions are shown in SEQ ID NO.18, SEQ ID NO.19 and SEQ ID NO.20 respectively.

[0012] In an optional embodiment, the nucleic acid sequences of the antigen complementary determining regions (CDRs) of the light chain variable regions are shown as SEQ ID NO.9, SEQ ID NO.10 and SEQ ID NO.11, respectively;

[0013] The nucleic acid sequences of the antigen complementary determining region (CDR) of the heavy chain variable region are shown in SEQ ID NO.12, SEQ ID NO.13 and SEQ ID NO.14 respectively.

[0014] In an optional embodiment, the amino acid sequence of the light chain variable region is shown in SEQ ID NO.3;

[0015] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO.4.

[0016] In an optional embodiment, the nucleic acid sequence of the light chain variable region is shown in SEQ ID NO.1;

[0017] The nucleic acid sequence of the heavy chain variable region is shown in SEQ ID NO.2.

[0018] In a second aspect, the present invention provides a nucleic acid molecule encoding the light chain and heavy chain of the fully human anti-Der p 1 full molecule IgG antibody as described in any one of the aforementioned embodiments.

[0019] In an optional embodiment, the amino acid sequence of the light chain is shown in SEQ ID NO.7;

[0020] The amino acid sequence of the heavy chain is shown in SEQ ID NO.8.

[0021] In an optional embodiment, the nucleic acid sequence of the light chain is shown in SEQ ID NO.5;

[0022] The nucleic acid sequence of the heavy chain is shown in SEQ ID NO.6.

[0023] As mentioned above, the nucleic acid molecules encode the light and heavy chains of the fully human anti-Der p 1 full molecule IgG antibody. These nucleic acid molecules (DNA sequences) contain the genetic information required to encode the antibody light and heavy chains. These sequences are designed to express functional antibody molecules in host cells. The use of nucleic acid molecules encoding fully human antibodies can avoid the immune response that may be caused by animal antibodies and improve the safety and effectiveness of treatment.

[0024] These nucleic acid molecules can be inserted into expression vectors through genetic engineering techniques and then transferred into host cells for expression.

[0025] In a third aspect, the present invention provides an expression vector comprising a nucleic acid molecule as described in any one of the aforementioned embodiments and an expression control sequence operably linked to the nucleic acid molecule.

[0026] In a fourth aspect, the present invention provides a biomaterial comprising an expression cassette of a nucleic acid molecule as described in the foregoing embodiments, an expression vector as described in the foregoing embodiments, and a recombinant cell line.

[0027] In a fifth aspect, the present invention provides a host cell transformed by the expression vector described in the above embodiment.

[0028] In a sixth aspect, the present invention provides a use of a fully human anti-Der p 1 whole molecule IgG antibody as described in any one of the aforementioned embodiments in the preparation of a therapeutic drug for activating human macrophages.

[0029] In a seventh aspect, the present invention provides a drug for activating the function of human macrophages, comprising the fully human anti-Der p 1 whole molecule IgG antibody as described in any one of the aforementioned embodiments.

[0030] The present invention provides a fully human anti-Der p 1 whole molecule IgG antibody and its application. The fully human anti-Der p 1 whole molecule IgG antibody is an antibody prepared by sequence alignment, inserting the gene sequence of the antibody into an expression vector using genetic engineering technology, and then transferring it into an expression host. The antibody can effectively recognize the HDM allergen component Der p 1 protein and activate human macrophages. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0032] Figure 1 This is a schematic diagram of the results of the enzyme-linked immunosorbent assay in Example 4 of the present application;

[0033] Figure 2 This is a schematic diagram of the electrophoresis of SDS-PAGE to detect the purity of 923 antibody in Example 4 of the present application;

[0034] Figure 3 This is a schematic diagram of the results of Western-blot detection in Example 4 of the present application;

[0035] Figure 4 Schematic diagram of the affinity curve of the Biacore T100 antibody affinity test in Example 4 of the present application (KD=1.583e-7);

[0036] Figure 5 This is a schematic diagram of the gene expression changes of THP-1 cells stimulated by 923 antibody detected by q-PCR in Example 5 of the present application;

[0037] Figure 6 This is a schematic diagram of the protein level changes in THP-1 cells stimulated by 923 antibody detected by ELISA in Example 5 of the present application;

[0038] Figure 7 This is a schematic diagram of the gene expression changes of THP-1 cells stimulated by the 923 antibody F(ab')2 fragment detected by q-PCR in Example 5 of the present application;

[0039] Figure 8 This is a schematic diagram of the protein level changes in THP-1 cells stimulated by the ELISA detection of the 923 antibody F(ab')2 fragment in Example 5 of the present application. DETAILED DESCRIPTION

[0040] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. If no specific conditions are specified in the examples, the conditions are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be obtained commercially.

[0041] The present application provides a fully human anti-Der p 1 whole molecule IgG antibody in an embodiment, wherein the fully human anti-Der p 1 whole molecule IgG antibody comprises a light chain variable region and a heavy chain variable region.

[0042] The amino acid sequences of the antigen complementary determining regions (CDRs) of the light chain variable regions are shown in SEQ ID NO. 15, SEQ ID NO. 16 and SEQ ID NO. 17. The amino acid sequences of the antigen complementary determining regions (CDRs) of the heavy chain variable regions are shown in SEQ ID NO. 18, SEQ ID NO. 19 and SEQ ID NO. 20.

[0043] As mentioned above, the antibodies are fully human and are obtained from human sources through human B cells or genetic engineering technology, which helps to reduce immune responses and side effects that may occur in clinical applications because the human body generally has a higher tolerance to fully human antibodies.

[0044] Antibodies are made up of two parts: a light chain and a heavy chain. Each chain contains a variable region (VL and VH) that is responsible for binding to a specific antigen (Der p 1). This structure enables the antibody to specifically recognize and bind to the target allergen.

[0045] CDR is the region of the antibody variable region that directly contacts the antigen and determines the specificity and affinity of the antibody. The amino acid sequences of the light chain and heavy chain CDR are defined, and these sequences are the key to the antibody's recognition and binding to Der p 1. The specific combination of these sequences gives the antibody its unique recognition properties.

[0046] In summary, the fully human anti-Der p 1 whole molecule IgG antibody is an antibody prepared by sequence alignment, inserting the gene sequence of the antibody into an expression vector using genetic engineering technology, and then transferring it into an expression host. The antibody can effectively recognize the HDM allergen component Der p 1 protein and activate human macrophages. This example provides the key features of the fully human anti-Der p 1 IgG antibody, which are crucial for the function, specificity and potential clinical application of the antibody.

[0047] In some embodiments, the nucleic acid sequences of the antigen complementary determining regions (CDRs) of the light chain variable region are shown in SEQ ID NO.9, SEQ ID NO.10 and SEQ ID NO.11, respectively. The nucleic acid sequences of the antigen complementary determining regions (CDRs) of the heavy chain variable region are shown in SEQ ID NO.12, SEQ ID NO.13 and SEQ ID NO.14, respectively.

[0048] In some embodiments, the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 3. The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 4.

[0049] In some embodiments, the nucleic acid sequence of the light chain variable region is shown as SEQ ID NO. 1. The nucleic acid sequence of the heavy chain variable region is shown as SEQ ID NO. 2.

[0050] In the examples of the present application, a nucleic acid molecule is provided, encoding the light chain and heavy chain of the fully human anti-Der p 1 full molecule IgG antibody as described in any one of the aforementioned embodiments.

[0051] In some embodiments, the amino acid sequence of the light chain is shown as SEQ ID NO.7; and the amino acid sequence of the heavy chain is shown as SEQ ID NO.8.

[0052] In some embodiments, the nucleic acid sequence of the light chain is shown as SEQ ID NO.5; and the nucleic acid sequence of the heavy chain is shown as SEQ ID NO.6.

[0053] An expression vector is provided in an embodiment of the present application, comprising a nucleic acid molecule as described in any of the aforementioned embodiments and an expression control sequence operably connected to the nucleic acid molecule.

[0054] The above vector contains nucleic acid molecules encoding the light and heavy chains of fully human anti-Der p 1 full-molecule IgG antibodies, as well as expression regulatory sequences operably linked to these nucleic acid molecules. This design enables the expression vector to transcribe and translate the encoded antibody light and heavy chain genes in host cells, thereby producing functional antibody molecules. The use of this expression vector can efficiently express fully human antibodies in host cells, reduce the immune response that may be caused by animal-derived antibodies, and improve the safety and effectiveness of treatment.

[0055] A biomaterial is provided in an embodiment of the present application, comprising an expression cassette of a nucleic acid molecule as described in the foregoing embodiment, an expression vector as described in the foregoing embodiment, and a recombinant cell line.

[0056] A host cell is provided in an embodiment of the present application, which is transformed with the expression vector described in the above embodiment.

[0057] The present application provides an application of a fully human anti-Der p 1 whole molecule IgG antibody as described in any one of the aforementioned embodiments in the preparation of a therapeutic drug for activating human macrophages.

[0058] The present application provides a drug for activating the function of human macrophages, comprising the fully human anti-Der p 1 whole molecule IgG antibody as described in any one of the aforementioned embodiments.

[0059] Example 1: HDM antibody sequence alignment and screening

[0060] In this example, the mouse / human antibodies to HDM allergen components reported in the past were summarized, the CDR3 regions of the light and heavy chains were annotated using IMGT, and the BCR sequences similar to the light and heavy chain CDR3s of antibodies to known HDM allergen components were searched from the database huARdb to screen out potential candidate antibody sequences.

[0061] Experimental methods:

[0062] By summarizing the mouse / human antibodies of HDM allergen components reported in the past, the CDR3 regions of the light and heavy chains were annotated through IMGT.

[0063] The BCR sequences similar to the light and heavy chain CDR3 of antibodies of known HDM allergen components were searched from the database huARdb for the next experiment.

[0064] Example 2: Preparation, expression and affinity purification of antibodies corresponding to the screening sequences

[0065] In this example, the antibody variable region sequences obtained by screening were loaded into an expression plasmid vector containing the antibody Fc segment sequence, and the antibody was expressed by transfecting 293F cells. Subsequently, a series of separation and purification steps were performed to obtain high-purity antibodies to verify the expression and purification efficiency of the antibody.

[0066] Experimental methods:

[0067] (1) Loading the screened variable region sequence onto an expression vector plasmid containing an antibody constant region gene for plasmid synthesis.

[0068] (2) Dissolve the synthesized plasmid with an appropriate amount of deionized water. Take 50 μL Fast-T1 competent cells and add 1 μL plasmid; let stand on ice for 30 minutes; heat shock in a 42°C water bath for 90 seconds; let stand on ice for 2 minutes. Add 950 μL culture medium to each tube of competent cells and place in a cell culture shaker at 37°C, 220 rpm for 1 hour.

[0069] (3) Take the reaction solution to transform competent bacteria, spread them on plates with corresponding resistance, and select clones for sequencing the next day. The clones with correct sequencing results are preserved and expanded to extract plasmids.

[0070] (4) Eukaryotic expression of whole molecule IgG:

[0071] A. Take 100 μg of the recombinant heavy chain plasmid and 100 μg of the light chain plasmid in 1 mL of Opti-MEM medium, which is solution A; take 400 μL of PEI transfection reagent in 0.6 mL of Opti-MEM medium, which is solution B. Add solution B to solution A and let stand at 37°C for 20 minutes.

[0072] B. Treat 293F cells. Centrifuge the 293F cells and resuspend them in 293F Expression Medium. Count the cells and calculate the cell viability ratio using trypan blue. Pipette 1.00×108 cells into a culture flask and make up to 98 mL with 293F Expression Medium.

[0073] C. After 20 minutes, add 2 mL of DNA and PEI transfection reagent complex to the prepared 293F cells.

[0074] D. Culture the cells in a shaking incubator at 8% CO2, 120 rpm, and 37°C. Collect the cell supernatant after 5 days.

[0075] (5) The collected cell culture supernatant was filtered through 0.45 μm and 0.22 μm filter membranes, and the equilibration solution and the eluate were filtered at the same time. The protein was purified using the AKAT purifier 100 protein purification system according to the standard steps of Protein A purification, with the sample loaded at a flow rate of 1 mL / min and the elution at a flow rate of 1.1 mL / min.

[0076] Example 3: Screening of fully human anti-HDM / Der p 1 whole molecule antibodies

[0077] In this example, the ELISA method was used to screen the expressed antibodies, and the antibodies that bind to HDM were selected to evaluate the specificity and affinity of the antibodies.

[0078] Experimental methods:

[0079] (1) Crude dust mite extract HDM was coated on a 96-well ELISA plate and diluted to 1 μg / mL with coating solution (0.1 M carbonate buffer, pH 9.6). 100 μL was added to each well and incubated at 4°C overnight.

[0080] (2) Wash with PBST washing solution (PBS containing 0.05% Tween) for 5 times, add 5% BSA (300 μL / well) for blocking, incubate at 37°C for 2 hours, wash 5 times and set aside.

[0081] (3) Dilute the antibody to 50 ng / mL, add 100 μL of antibody to each well, incubate at 37°C for 1 hour, and wash 5 times with PBST.

[0082] (4) 100 μL / well of goat anti-human Ig-HRP secondary antibody (Thermo) diluted 1:20,000 was added to the wells, incubated at 37°C for 1 hour, and washed 5 times with PBST.

[0083] (5) Add 100 μL / well of peroxidase substrate colorimetric solution, and terminate the reaction with 2M sulfuric acid after 7 minutes at room temperature. Detection was performed using a full-wavelength microplate reader (Thermolabsystems, USA), with a dual wavelength of 450nm / 5700nm for colorimetry. The antibody with a detection result greater than 2.5 times the OD value of the blank well was the antibody that bound to HDM. Finally, one antibody was selected and named 923.

[0084] Example 4: Characterization of anti-HDM / Der p 1 whole molecule IgG

[0085] In this example, the characteristics of the screened 923 antibodies were analyzed by enzyme-linked immunosorbent assay, SDS-PAGE, Western-blot and affinity detection to comprehensively evaluate their interaction with Der p 1 and biological functions.

[0086] Experimental methods:

[0087] (1) ELISA:

[0088] A. The dust mite allergy component Der p 1 was coated on a 96-well ELISA plate and diluted to 1 μg / mL with coating solution (0.1 M carbonate buffer, pH 9.6). 100 μL was added to each well and incubated at 4°C overnight.

[0089] B. Wash with PBST washing solution (PBS containing 0.05% Tween) for 5 times, add 5% BSA (300 μL / well) for blocking, incubate at 37°C for 2 hours, wash 5 times and set aside.

[0090] C. Add 100 μL of anti-Der p 1 whole molecule IgG (2 μg / mL) to each well and incubate at 37°C for 2 hours.

[0091] D. Add 100 μL of goat anti-human secondary antibody (1:20,000 dilution) to each well and incubate at 37°C for 1 hour.

[0092] E. Wash 5 times with PBST.

[0093] F. Add peroxidase substrate colorimetric solution, terminate the reaction with 2M sulfuric acid after 7 minutes at room temperature, and detect the protein absorbance value on a microscope.

[0094] The results are as follows Figure 1 As shown, the 923 antibody can react significantly with the recombinant Der p 1 protein.

[0095] (2) SDS-PAGE detection of purified 923 antibody:

[0096] A. Prepare 8% polyacrylamide gel.

[0097] B. Add 5× loading buffer to the supernatant of untransfected 293F cells, the supernatant of 293F cells before purification, the ultrafiltered 923 antibody, the flow-through collected during purification, the washing solution, and the control human IgG, respectively, mix well, and place in a metal bath at 100°C for 10 minutes. Prepare a non-reduced, unboiled sample for each of the ultrafiltered 923 antibody and the control human IgG.

[0098] C. Perform electrophoresis after adding the sample. The voltage of the concentrated gel is 80V, and that of the separation gel is 110V. The electrophoresis can be terminated when the bromophenol blue runs out.

[0099] D. Coomassie blue staining gel for 30 minutes, deionized water decolorization until the background is clean and the target band is clearly visible ( Figure 2 ).

[0100] (3) Western-blot:

[0101] A. Add 5× loading buffer to crude dust mite extract HDM, Der p 1 recombinant protein and negative control bovine serum albumin BSA, mix well and place in a 100℃ metal bath for 10 minutes.

[0102] B. Perform 12% SDS-PAGE electrophoresis.

[0103] C. Transfer the protein to nitrocellulose membrane by electroporation.

[0104] D. The membrane was quickly blocked for 30 minutes and then incubated with 0.4 μg / mL 923 antibody at 4°C overnight.

[0105] E. After washing with TBST, incubate with 1:10000 diluted HRP-goat anti-human IgG for 1 hour.

[0106] F, and ECL luminescence kit were exposed to the gel imaging system, and the results were shown in Figure 3 .

[0107] (4) Affinity detection

[0108] The coupling conditions were optimized according to the isoelectric point and the protocol of BiacoreX100 control soft, and sodium acetate was selected as the coupling dilution buffer for slope optimization.

[0109] A. Dilute the 923 antibody sample to 25 μg / mL with this buffer and couple it to a CM5 chip (GE#BR100012).

[0110] B. Default coupling level 1500RU.

[0111] C. Dilute the Der p 1 series samples with pH 7.4 Running buffer to a series of concentrations of 0 μg / mL, 15.625 μg / mL, 31.25 μg / mL, 62.5 μg / mL, 125 μg / mL, and 250 μg / mL.

[0112] D. Set the injection time to 180 s, the dissociation time to 10 min, and the regeneration buffer to 50 mM pH 2.2 Gly-HCl.

[0113] E. Perform the test on the BiacoreX100 control soft protocol. The affinity KD of the antibody tested was 1.583e(-7) (see Figure 4 ).

[0114] Example 5: Detection of the activation effect of anti-Der p 1 whole molecule IgG on human macrophages

[0115] In this example, the effect of 923 antibody on the release of IL-1β by human macrophage THP-1 was detected to evaluate its immunomodulatory function and potential therapeutic effect.

[0116] Experimental methods:

[0117] (1) Antibody 923 stimulates human macrophage THP-1 to release IL-1β:

[0118] A. THP-1 cells were seeded in 24-well plates, with 3×10 5 Cells were cultured and induced to differentiate by adding 10 ng / mL PMA for 48 hours, and then the culture medium was replaced with fresh medium.

[0119] B. 0.5 μg / mL of 923 antibody and 0.5 μg / mL of hIgG isotype control were added to the induced THP-1 cells and stimulated for 12 hours.

[0120] C. Collect the culture supernatant and use an RNA extraction kit and a cDNA synthesis kit to extract cDNA from the induced THP-1 cells.

[0121] D. Prepare the q-PCR reaction system. Add 5 μL SYBR Green, 3 μL enzyme-free water, 0.5 μL forward primer, 0.5 μL reverse primer, and 1 μL cDNA to each well. Perform q-PCR reaction. The results are shown in Figure 5 .

[0122] E. Use Human IL-1β ELISA kit to detect the IL-1β content in the cell supernatant. The results are shown in Figure 6.

[0123] (2) 923 antibody F(ab')2 fragment stimulates human macrophage THP-1 to release IL-1β:

[0124] A. The F(ab')2 fragment was prepared from the 923 antibody using a F(ab')2 preparation kit, and the prepared F(ab')2 fragment was detected by SDS-PAGE electrophoresis.

[0125] B. THP-1 cells were seeded in 24-well plates, with 3×10 5 Cells were cultured and induced to differentiate by adding 10 ng / mL PMA for 48 hours, and then the culture medium was replaced with fresh medium.

[0126] C. 50 μg / mL of 923 antibody F(ab')2 was added to the induced THP-1 cells and stimulated for 12 hours.

[0127] D. Collect the culture supernatant and use an RNA extraction kit and a cDNA synthesis kit to extract cDNA from the induced THP-1 cells.

[0128] E. Prepare the q-PCR reaction system. Add 5 μL SYBR Green, 3 μL enzyme-free water, 0.5 μL forward primer, 0.5 μL reverse primer, and 1 μL cDNA to each well. Perform q-PCR reaction. The results are shown in Figure 7 .

[0129] F. Use Human IL-1β ELISA kit to detect the IL-1β content in the cell supernatant. The results are shown in Figure 8 .

[0130] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A fully human anti-Der p 1 whole molecule IgG antibody, characterized in that: The fully human anti-Der p 1 whole molecule IgG antibody comprises a light chain variable region and a heavy chain variable region; The amino acid sequences of the antigen complementary determining regions (CDRs) of the light chain variable regions are shown in SEQ ID NO.15, SEQ ID NO.16 and SEQ ID NO.17 respectively; The amino acid sequences of the antigen complementary determining regions (CDRs) of the heavy chain variable regions are shown in SEQ ID NO.18, SEQ ID NO.19 and SEQ ID NO.20 respectively.

2. The fully human anti-Der p 1 whole molecule IgG antibody according to claim 1, characterized in that: The nucleic acid sequences of the antigen complementary determining regions (CDRs) of the light chain variable regions are shown in SEQ ID NO.9, SEQ ID NO.10 and SEQ ID NO.11 respectively; The nucleic acid sequences of the antigen complementary determining region (CDR) of the heavy chain variable region are shown in SEQ ID NO.12, SEQ ID NO.13 and SEQ ID NO.14 respectively.

3. The fully human anti-Der p 1 whole molecule IgG antibody according to claim 1, characterized in that: The amino acid sequence of the light chain variable region is shown in SEQ ID NO.3; The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO.

4.

4. The fully human anti-Der p 1 whole molecule IgG antibody according to claim 1, characterized in that: The nucleic acid sequence of the light chain variable region is shown in SEQ ID NO.1; The nucleic acid sequence of the heavy chain variable region is shown in SEQ ID NO.

2.

5. A nucleic acid molecule, characterized in that Encoding the light chain and heavy chain of the fully human anti-Der p 1 full-molecule IgG antibody according to any one of claims 1 to 4; Preferably, the amino acid sequence of the light chain is shown as SEQ ID NO.7; the amino acid sequence of the heavy chain is shown as SEQ ID NO.8; Preferably, the nucleic acid sequence of the light chain is shown as SEQ ID NO.5; and the nucleic acid sequence of the heavy chain is shown as SEQ ID NO.

6.

6. An expression vector, characterized in that: The method comprises the nucleic acid molecule as claimed in claim 5 and an expression control sequence operably linked to the nucleic acid molecule.

7. A biomaterial, characterized in that: An expression cassette comprising the nucleic acid molecule of claim 5, an expression vector and a recombinant cell line of claim 6.

8. A host cell, characterized in that Transformed with the expression vector according to claim 6.

9. Use of the fully human anti-Der p 1 whole molecule IgG antibody according to any one of claims 1 to 4 in the preparation of a therapeutic drug for activating human macrophages.

10. A drug for activating human macrophage function, characterized in that: The invention comprises the fully human anti-Der p 1 whole molecule IgG antibody as described in any one of claims 1 to 4.

Images