39 results about "Mite allergen" patented technology

The invention provides a method for producing a modified major mite allergen, obtained by altering a major allergen of house dust mites by gene engineering, which involve the steps of culturing prokaryotic or eukaryotic host cells transformed with a replication vector containing a gene encoding modified major mite allergen expressed from a promoter and collecting the expressed modified major mite allergen from the culture. The present invention provides a modified major mite allergen produced by the method, DNA molecules encoding same, and a pharmaceutical composition containing the modified major mite allergen.

The invention provides a novel lyophilized mite allergen vaccine and a preparation method thereof. The vaccine is prepared by mixing and lyophilizing mite allergen activated protein, a lyophilization protective agent and phosphate buffer solution with pH value of 7.0-8.5 based on the weight ratio of 1:(100-1000):(70-140). Compared with the traditional water preparation, glycerol preparation and preparation containing an aluminum hydroxide adjuvant of the mite allergen vaccine, the lyophilized mite allergen vaccine overcomes use inconvenience of the glycerol preparation, instability of the water preparation and side effects resulting from the preparation containing the aluminum hydroxide adjuvant, thus providing an efficient vaccine product with good stability and good safety for patients allergic to dust mites.

The invention discloses a method for detecting mite allergen specific antibody in blood serum. The method comprises the following steps of: coupling mite allergen with surface carboxyl modified magnetic microspheres to prepare immunomagnetic microspheres; and mixing and incubating the blood serum to be detected and the immunomagnetic microspheres in holes of an elisa (enzyme-linked immunosorbent assay) plate to combine mite allergen with mite allergen specificity IgE in the blood serum, and detecting by adopting an enzyme-linked immunosorbent method after the elisa plate is washed. In the method disclosed by the invention, a mite allergen is directly coupled onto the surfaces of magnetic microspheres, thus all the impurities except the mite allergen specific antibody in the blood serum can be conveniently removed; and targeted mite allergen specificity antibody detection is carried out, detection time is shortened, and sensitivity, specificity and accuracy of detection are improved. The mite allergen immunomagnetic microspheres prepared by the invention can be used in standard automatic detection to improve detection efficiency.

The invention provides a method for preparing an allergen diagnostic reagent of acute allergic diseases caused by dermatophagoides pteronyssinus (Der p for short). The allergen diagnostic reagent is Der p allergen pricking liquid. The invention also provides a method for preparing an allergen diagnostic reagent raw material, namely Der p pure mite allergen, a method for evaluating the valence of the allergen diagnostic reagent and a purpose for specifically diagnosing the allergic diseases. The allergen diagnostic reagent achieves the aim for detecting whether a patient is allergic to a matter to cause a disease by a sensitized principle. The Der p pure mite allergen raw material has short culture period and high purity, and the pricking liquid made of the raw material has high activity, thereby greatly reducing a skin test risk of the allergic patient and improving the safety of clinical medical treatment.

The invention relates to recombinant lactobacillus, use of the same and a pharmaceutical composition including the same. The recombinant lactobacillus includes a heterologous nucleic acid sequence. The heterologous nucleic acid sequence encodes at least an immunogenic fragment of the mite allergens Der p 1, Der p 2, or Blo t 5, or an immunogenic homolog thereof. A respective fragment of Der p 1 includes at least 8% of the amino acid sequence of the mite allergen. A method of modulating the immune response to an allergen in a mammal as well as a pharmaceutical composition and a kit are also disclosed.

The invention relates to a pharmaceutical product comprising an allergen extract or an allergoid thereof for the treatment and / or prevention of allergy and allergic asthma caused by house dust mites, which extract comprises at least one extract of mite bodies selected from the following groups a)-b): a) An extract of Der p mite bodies, and b) An extract of Der f mite bodies, and at least one extract of mite cultures selected from the following groups c)-g): c) An extract of Der p faecal particles, d) An extract of Der f faecal particles, e) An extract of Der f whole mite culture, f) An extract of an Der p whole mite culture, and g) a combination of extracts c) to f).

The present invention refers to recombinant ADN molecules coding to hybrids polypeptides of different allergens from D. pteronyssinus useful for the prevention and treatment of allergies, particularly allergies caused by mites. Specifically, the invention describes hybrid proteins composed of fragments of allergens Derp p l y Derp p 2 with hypoallergenic characteristics and maintain their immunogenic capacity, being particularly useful for the treatment of allergy. The invention also describes the production methods of these polypeptides in heterologous expression systems. Besides, the invention describes efficient purification methods of these hybrid proteins.

The invention relates to recombinant lactobacillus, use of the same and a pharmaceutical composition including the same. The recombinant lactobacillus includes a heterologous nucleic acid sequence. The heterologous nucleic acid sequence encodes at least an immunogenic fragment of the mite allergens Der p 1, Der p 2, or Blo t 5, or an immunogenic homolog thereof. A respective fragment of Der p 1 includes at least 8 % of the amino acid sequence of the mite allergen. A method of modulating the immune response to an allergen in a mammal as well as a pharmaceutical composition and a kit are also disclosed.

The present invention comprises a method, materials and apparatus for investigating asthma in humans using dust mite allergen. The dust mite allergen is prepared to be of a controlled size such that particles are respirable, effectively aerosolized, and deliver a significant amount of allergen. The dust mite allergen is applied in a controlled manner within an environmental exposure chamber to elucidate the etiological links between dust mite allergen concentration and asthma response in humans. The environmental exposure chamber is specially-designed to promote homogeneity of allergen concentration. Preferably, the dust mite allergen preparation comprises particles having an average diameter of less than 25 microns, and more preferably 5-10 microns. Correlation between aerosolized particle count and allergen concentration enables “real-time” allergen concentration estimates without the use of expensive and time-consuming assay techniques.

The invention provides a monoclonal antibody capable of recognizing an allergen of DerP2, wherein the monoclonal antibody is secreted by a hybridoma cell strain DerP2 with the accession number being CCTCC NO:C2014112. The invention further provides an ELISA kit for quantitative detection of dermatophagoides pteronyssinus allergens of DerP2 and Derf2. The ELISA kit includes an enzyme label plate, a specimen diluent, a coating antibody, a detecting antibody, a detecting antibody diluents, a standard substance, a washing liquid, a substrate chromogenic reagent, and a stopping solution, wherein the coating antibody is the monoclonal antibody related in the claim 1, and the detecting antibody is an anti-DerP2 polyclonal antibody marked with HRP. The anti-DerP2 monoclonal antibody is excellent in specificity and produces no cross reaction with a DerP1 protein. Therefore, based on the anti-DerP2 monoclonal antibody, the fast, specific, and sensitive in-vitro diagnostic kit for detecting the allergens is established.

A safe and efficient recombinant mite allergen is provided as a therapeutic agent or a diagnostic agent for mite allergic diseases, which contains no anaphylaxis-inducing impurities. The following recombinant protein (a) or (b) is provided: (a) a protein comprising the amino acid sequence represented by SEQ ID NO: 2 or 35; or (b) a protein comprising an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 2 or 35 by deletion, substitution, or addition of one or several amino acids and having mite allergen activity.

A safe and efficient recombinant mite allergen is provided as a therapeutic agent or a diagnostic agent for mite allergic diseases, which contains no anaphylaxis-inducing impurities. The following recombinant protein (a) or (b) is provided:(a) a protein comprising the amino acid sequence represented by SEQ ID NO: 2 or 35; or(b) a protein comprising an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 2 or 35 by deletion, substitution, or addition of one or several amino acids and having mite allergen activity.

The invention relates to a recombinant mite allergen protein composition, the recombinant mite allergen protein composition comprises recombinant mite I-type allergens DP1 and DF1 protein and recombinant mite II-type allergens DP2 and DF2 protein which are respectively mixed according to the activity ratio of 1: 1: 1: 1 or mixed according to the mass ratio of (1-5): (1-20): (1-10): (1-20). Compared with an existing product, the protein composition has the advantages that (1) the recombinant allergen has high purity, controllable quality and the like; (2) the recombinant protein has good specificity; and (3) compared with a natural extracting solution, the protein composition has better immunogenicity, the risk of serious allergic side reactions is reduced, and the desensitization treatmenteffect is improved; secondly, the recombinant allergen composition disclosed by the invention not only has better activity than natural allergen protein in the same proportion, but also has activityhigher than that of a corresponding natural single component compared with that of a single component, and has a product development and application prospect.

The invention relates to a pharmaceutical product comprising an allergen extract or an allergoid thereof for the treatment and / or prevention of allergy and allergic asthma caused by house dust mites, which extract comprises at least one extract of mite bodies selected from the following groups a)-b): a) An extract of Der p mite bodies, and b) An extract of Der f mite bodies, and at least one extract of mite cultures selected from the following groups c)-g): c) An extract of Der p faecal particles, d) An extract of Der f faecal particles, e) An extract of Der f whole mite culture, f) An extract of an Der p whole mite culture, and g) a combination of extracts c) to f).

The invention provides an intelligent mite cultivating and collecting device, which comprises a box body with a sealed holding space and a feeding device 2, a collecting device, a lighting device 4, atemperature control device 5 and a humidity control device 6 which are arranged in the sealed holding space. The invention further provides a method for standardly cultivating the mite by using the device. The method is time-saving and labor-saving, able to realize the scaled and standardized mite cultivation, and good for pushing the development and study of the mite allergen and vaccine.

The invention discloses a moisture-proof mite allergen soluble microneedle. The moisture-proof mite allergen soluble microneedle comprises a needle body and a backing, wherein the needle body comprises mite allergen and a composite matrix material, the composite matrix material is a mixture of a methyl vinyl ether-maleic anhydride copolymer and polyvinyl alcohol, and the content ratio of the methyl vinyl ether-maleic anhydride copolymer to the polyvinyl alcohol is (1:1.5)-(2:1). According to the moisture-proof mite allergen soluble microneedle, by screening the appropriate matrix material type and content (Gantrez S-97+PVA) and the appropriate drug loading ratio, the prepared mite allergen soluble microneedle can still have good mechanical strength under a high-humidity condition, is easy to pierce into the skin for drug administration, and does not affect the activity of the mite allergen.

The invention provides an intelligent mite cultivating and collecting device, which comprises a box body with a sealed holding space and a feeding device 2, a collecting device, a lighting device 4, atemperature control device 5 and a humidity control device 6 which are arranged in the sealed holding space. The invention further provides a method for standardly cultivating the mite by using the device. The method is time-saving and labor-saving, able to realize the scaled and standardized mite cultivation, and good for pushing the development and study of the mite allergen and vaccine.

The present invention relates to recombinant DNA encoding hybrid polypeptides of different allergens from the European dust mite useful for the treatment and prevention of allergies, more particularly those caused by mites. Specifically, the present invention describes a hybrid protein composed of fragments of the allergens Derp p 1 y Derp p 2, which has low allergenicity and maintains immunogenicity, and is particularly useful for the treatment of allergies. The invention also describes methods for producing these polypeptides in heterologous expression systems. Furthermore, the present invention describes efficient purification methods for these hybrid proteins.

The invention relates to a hybridoma cell strain capable of secreting a monoclonal antibody of a seventeenth component of an anaphylactic acid mite allergen and application of the hybridoma cell strain, and belongs to the technical field of immunodetection. According to the hybridoma cell strain, the hybridoma cell strain secreting the monoclonal antibody of the seventeenth component of the achnoid tropicalis allergen is obtained through screening for the first time, the monoclonal antibody secreted by the hybridoma cell strain has good sensitivity to Blo t 17, EC50 is 26.71 ng / mL, and therefore low-concentration Blo t 17 can be accurately detected. The monoclonal antibody disclosed by the invention is applied to a Blo t 17 detection product, and high-efficiency detection of low-concentration Blo t 17 can be realized.

The invention discloses a sublingual temperature-sensitive gel containing mite allergen. The sublingual temperature-sensitive gel is composed of mite allergen, a cationic monomer and a temperature-sensitive gel material, wherein the content ratio of the cationic monomer to the temperature-sensitive gel material is (1:2)-(1:1). According to the sublingual temperature-sensitive gel containing mite allergen, the sublingual temperature-sensitive gel is provided on the basis of a common sublingual drop containing mite allergen, the advantage of convenience in administration of the sublingual drop is maintained, the retention time of a vaccine under the tongue is prolonged, and the drug effect is improved; the gelling time of the temperature-sensitive gel is shortened, the loss caused by swallowing in the administration process is reduced, and the dosage is reduced; and the thermo-sensitivity of the temperature-sensitive gel to adsorb protein is overcome, the adsorption of the gel to the protein in a liquid state is improved, and then the dosage is reduced.

The invention discloses an adjuvant-containing mite allergen soluble microneedle patch. The adjuvant-containing mite allergen soluble microneedle patch comprises a needle body and a backing, wherein the needle body comprises a mite allergen, a composite adjuvant and a matrix material; the composite adjuvant is a mixture of microcrystalline tyrosine and alfalfa polysaccharide; and the content ratio of the microcrystalline tyrosine to the alfalfa polysaccharide is (2:1)-(1:2). According to the mite allergen soluble microneedle patch provided by the invention, by selecting the appropriate type and content of the composite adjuvant and the mass ratio of the matrix material in the needle body, the prepared mite allergen soluble microneedle patch is appropriate in mechanical strength, good in treatment effect and excellent in safety.

The present invention provides a modified Dermatophagoides pteronyssinus allergen Der p 5 protein which has ability to inhibit IgE binding when exposed against to the antigen. A method for treating allergy comprising administrating a therapeutically effective dose of the modified D. pteronyssinus allergen Der p 5 protein to a subject suffering from allergy Der p 5 is also provided.

A method for purifying a modified major mite allergen obtained by the genetic recombination technique and a purified modified major mite allergen obtained by said method for purification are provided. A method for purifying a modified major mite allergen obtained by the genetic recombination technique, which comprises the purification steps: (1) Washing and recovering inclusion bodies containing a modified major mite allergen obtained by the genetic recombination technique with MF membrane; (2) Dissolving said inclusion bodies followed by refolding; (3) Concentrating a solution containing the modified major mite allergen with simultaneous removal of low molecular weight components with ultrafiltration membrane; (4) Recovering the modified major mite allergen in non-adsorbed fractions with an anion exchanger; (5) Recovering the modified major mite allergen in adsorbed fractions with a hydrophobic gel; and (6) Recovering the modified major mite allergen in adsorbed fractions with an anion exchanger, and a modified major mite allergen with high purity obtained by said method for purification.

The invention relates to a mite allergen specific yolk immunoglobulin antibody and an anti-allergic preparation thereof. The mite allergen specific yolk immunoglobulin antibody is mainly prepared by the following method: extracting a mite allergen, carrying out supplementary immunization on chickens, preparing crude extract, finely extracting and purifying. The mite allergen specific yolk immunoglobulin antibody and the preparation thereof, provided by the invention, have the advantages of high antibody activity and specific defending effect on local sensitization caused by mites, and have a good rejection effect on the local sensitization caused by sensibiligen; the mite allergen specific yolk immunoglobulin antibody has no immunogenicity, no adverse effect on skin mucous membranes of human bodies and high safety performance.

The invention relates to a monoclonal antibody of a 37-th component of an anaphylactogen of chaenomeles tropicalis, a hybridoma cell strain and application thereof, and belongs to the technical field of immunodetection. The hybridoma cell strain secreting the monoclonal antibody of the 37th component of the achynx tropicalis allergen is obtained through screening for the first time, the monoclonal antibody secreted by the hybridoma cell strain has good sensitivity to Blo t 37, EC50 is 27.56 ng / mL, and therefore low-concentration Blo t 37 can be accurately detected. The monoclonal antibody disclosed by the invention is applied to a Blo t 37 detection product, and efficient detection of low-concentration Blo t 37 can be realized.

The invention discloses a slow-release soluble microneedle delivery system for mite-allergen-loaded microspheres. The system comprises a needle body and a backing, wherein the needle body comprises the mite allergen microspheres and a matrix material, the matrix material is a mixture of a methyl vinyl ether-maleic anhydride copolymer and polyethylene glycol, and the content ratio of the methyl vinyl ether-maleic anhydride copolymer to the polyethylene glycol is (2:1)-(1:1). According to the system, the specific component types and content ratio of the mite allergen microspheres are selected, so that the prepared mite allergen slow-release microspheres are excellent in performance and can well load mite allergen; and the specific component types and content ratio of the matrix material and the specific drug loading ratio are selected, so that the prepared mite allergen slow-release soluble microneedle is excellent in formability, mechanical strength and other performance, the release time of the drug is greatly prolonged, the treatment effect is superior to that of a non-slow-release mite allergen soluble microneedle, the administration frequency of the mite allergen can be reduced, and the compliance of a patient can be improved.

The invention provides modified hydroxyapatite for adsorbing dermatophagoides pteronyssinus allergens as well as a synthesis method and application of the modified hydroxyapatite. Dermatophagoides pteronyssinus allergen in the air is mainly suspended in the form of protein, and when exposed in the environment, a human body passively inhales or contacts with the dermatophagoides pteronyssinus allergen to induce allergic rhinitis, asthma and other diseases. The modified hydroxyapatite has a unique porous crystal structure, the content of the main component Ca10 (PO4) 6 (OH) 2 is larger than or equal to 99 wt%, the surface of the modified hydroxyapatite is rich in hydroxyl (-OH), functional groups such as amino (-NH2) and carboxyl (-COOH) can be introduced through targeted modification, and the modified hydroxyapatite is in a sheet-shaped or rod-shaped crystal shape. The dust mite allergens in air are removed through the synergistic effect of physical adsorption and chemical adsorption, and the removal rate of the dust mite allergens Der p 1 and Der f 1 is increased under the room temperature condition. The invention further discloses a synthesis method of the modified hydroxyapatite and application of the modified hydroxyapatite in the field of air purification, and an efficient solution is provided for indoor allergen prevention and control.

The invention provides a dust mite allergen quick-disintegrating sublingual tablet vaccine and a preparation method thereof. The vaccine is a quick-disintegrating sublingual tablet of dust mite allergens prepared by direct freeze-drying method, and contains dust mite allergens and corresponding excipients, flavoring agent, pigments, etc. in which the dust mite allergens are Dermatophagoides pteronyssinus allergens or Dermatophagoides farinae allergens or mixed dust mite allergens of both. The sublingual tablet vaccine has the advantages of stable biological activity, long effect period, easy storage, convenient use, disintegration, quick release of active ingredients and the like compared with the traditional allergen sublingual drop vaccine and can provide more convenient and reliable specific desensitization treatment means for patients allergic to dust mites.