# US 8,465,945 B2 · Method for production and use of mite group 1 proteins > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,465,945 B2 - Document ID: US8465945B2 - Status: Granted - Kind: B2 - Application number: 11/542,659 - Filed: 2006-10-02 - Published: 2007-04-12 - Granted: 2013-06-18 - Earliest claimed: 1999-10-15 - Assignees: Merck Patent GmbH - Applicants: Elaine A. Best; Martin J. McDermott - Inventors: Elaine A. Best; Martin J. McDermott - Web page: https://patentgrep.com/patent/US8465945B2 ## CPC - C07K 16/18: against material from animals or humans - A61P 37/00: Drugs for immunological or allergic disorders - C07K 14/43531: {from mites} - A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) ## Abstract The present invention includes a method to produce a recombinant mite Group 1 protein in a methyltrophic yeast or an Escherichia coli microorganism. The present invention also relates to a recombinant mite Group 1 protein obtained by such a method, such a recombinant protein being able to selectively bind IgE or cause proliferation of a T cell that proliferate in response to a native mite Group 1 protein. Also included in the present invention is the use of such a recombinant mite Group 1 protein to detect mite allergy or to reduce an allergic response to a mite Group 1 protein. The present invention also includes novel mite Group 1 nucleic acid molecules, proteins, recombinant molecules, and recombinant cells, as well as uses thereof. ## Claims ### Claim 1 A method to produce a recombinant protein comprising a recombinant mite Group 1 protein, comprising the steps of: (a) culturing a methyltrophic yeast microorganism transformed with a nucleic acid molecule encoding said recombinant mite Group 1 protein; and (b) recovering said recombinant mite Group 1 protein from said methyltrophic yeast microorganism; wherein said recombinant mite Group 1 protein is a Dermatophagoides pteronyssinus, recombinant mite Group 1 protein (Der p 1) and said nucleic acid molecule consists of the nucleic acid sequence of SEQ ID NO:10, and wherein said nucleic acid molecule is optionally fused to a signal sequence. ### Claim 2 (depends on 1) The method of claim 1, wherein said methyltrophic yeast microorganism is of a genus selected from the group consisting of a Pichia microorganism, a Hansenula microorganism, a Torulopsis microorganism, and a Candida microorganism. ### Claim 3 (depends on 1) The method of claim 1, wherein said methyltrophic yeast microorganism is of a species selected from the group consisting of Pichia pastoris, Pichia acaciae, Pichia anomala, Pichia augusta, Pichia capsulata, Pichia fabianii, Pichia farinosa, Pichia guilliermondii, Pichia methanolica, Pichia norvegensis, Pichia pinus, Pichia stipitis, and Hansenula polymorpha. ### Claim 4 (depends on 1) The method of claim 1, wherein said methyltrophic yeast microorganism is Pichia pastoris. ### Claim 5 (depends on 1) The method of claim 1, wherein said nucleic acid molecule encodes a recombinant mite Group 1 protein selected from the group consisting of a pro-form of a recombinant mite Group 1 protein and a mature form of a recombinant mite Group 1 protein. ### Claim 6 (depends on 1) The method of claim 1, wherein said nucleic acid molecule encodes a recombinant mite Group 1 protein fused to a S. cerevisiae alpha factor signal sequence. ### Claim 7 (depends on 1) The method of claim 1, wherein said recombinant mite Group 1 protein is a recombinant mite Group 1 protein consisting of the amino acid sequence of SEQ ID NO:11. ### Claim 8 (depends on 1) The method of claim 1, wherein said recombinant mite Group 1 protein is selected from the group consisting of a pro-form of said recombinant mite Group 1 protein and a mature form of said recombinant mite Group I protein. ### Claim 9 (depends on 1) The method of claim 1, wherein said recombinant mite Group 1 protein is secreted by said methyltrophic yeast microorganism. ### Claim 10 A methyltrophic yeast microorganism comprising a nucleic acid molecule encoding a mite Group 1 protein wherein, said nucleic acid molecule consists of the nucleic acid sequence of SEQ ID NO:10, and said recombinant mite Group 1 protein is a Dermatophagoides pteronyssinus, recombinant mite Group 1 protein (Der p 1), said nucleic acid molecule being operatively linked to a transcription control sequence and wherein sad nucleic acid molecule is optionally fused to a signal sequence. ### Claim 11 (depends on 10) The methyltrophic yeast microorganism of claim 10, wherein said recombinant mite Group 1 protein is a recombinant mite Group 1 protein consisting of the amino acid sequence of SEQ ID NO:11. ### Claim 12 (depends on 10) The methyltrophic yeast microorganism of claim 10, wherein said methyltrophic yeast microorganism is of a genus selected from the group consisting of a Pichia microorganism, a Hansenula microorganism, a Torulopsis microorganism, and, a Candida microorganism. ### Claim 13 (depends on 12) The methyltrophic yeast microorganism of claim 12, wherein aid Pichia microorganism is P. pastoris. ### Claim 14 (depends on 13) The methyltrophic yeast microorganism of claim 13, wherein said nucleic acid molecule encodes a recombinant mite Group 1 protein fused to a S. cerevisiae alpha factor signal sequence.