RECOMBINANT DERMATOPHAGOIDES PTERONYSSINUS TYPE 2 ALLERGEN PROTEIN AND ITS PREPARATION METHOD AND APPLICATION

• US 20200131235A1
• Filed: 12/28/2017
• Published: 04/30/2020
• Est. Priority Date: 12/31/2016
• Status: Active Grant

• Current Assignee: Zonhon Biopharma Institute Inc.
• Original Assignee: Zonhon Biopharma Institute Inc.
• Inventors: MA, Bruce Yong, FAN, Yu, WANG, Anliang, WANG, Jun
• Granted Patent: US 11,319,353 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 38/17 from animals; from humans enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 39/35 Allergens
• CPC Class Codes: A61P 11/02 Nasal agents, e.g. decongestants
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: C07K 1/18 Ion-exchange chromatography
• CPC Class Codes: C07K 1/20 Partition-, reverse-phase or hydrophobic interaction chromatography
• CPC Class Codes: C07K 1/30 by precipitation
• CPC Class Codes: C07K 1/36 by a combination of two or more processes of different types
• CPC Class Codes: C07K 14/43531 from mites
• CPC Class Codes: C12N 15/81 for yeasts
• CPC Class Codes: C12N 15/815 for yeasts other than Saccharomyces

Abstract

Provided are a recombinant group 2 allergen protein from Dermatophagoides pteronyssinus, and an encoding gene and preparation method thereof. The encoding gene is obtained by preforming codon optimization on a wild-type encoding gene of a group 2 allergen protein from Dermatophagoides pteronyssinus and is beneficial to expression in Pichia pastoris. Further, an acting element increasing target protein expression is added to the expression system.

Claims

1. A DNA sequence encoding Der p2 protein, having a base sequence as shown in SEQ ID NO:

1.

9. A DNA sequence of claim 1, wherein the DNA sequence is comprised in a vector pAO815, pPIC9, pPIC9K, pPIC3.5, pPIC3.5K, pPICZα

A, B, C or pGAPZ A, B, C.

10. A DNA sequence of claim 9, wherein the vector is comprised in Pichia pastoris strain SMD1168, GS115, KM71, X33 or KM71H.

11. A DNA sequence of claim 10, wherein there is a 242 bp interval between the DNA sequence encoding Der p2 protein and ATG of AOX1 on Pichia pastoris;

and the DNA sequence encoding the Der p2 protein is preceded by Kozak sequence GCCACCATGG.

2. A recombinant Der p2 protein, having an amino acid sequence as shown in SEQ ID NO:

3.

12. Recombinant Der p2 protein of claim 2, wherein the recombinant protein is encoded by the base sequence as shown in SEQ ID NO:

1.

13. Recombinant Der p2 protein of claim 12, wherein the DNA sequence is comprised in a vector pAO815, pPIC9, pPIC9K, pPIC3.5, pPIC3.5K, pPICZα

A, B, C or pGAPZ A, B, C.

14. The recombinant Der p2 protein of claim 13, wherein the vector is comprised in Pichia pastoris strain SMD1168, GS115, KM71, X33 or KM71H.

15. The recombinant Der p2 protein of claim 14, wherein there is a 242 bp interval between the DNA sequence encoding Der p2 protein and ATG of AOX1 on Pichia pastoris;

and the DNA sequence encoding the Der p2 protein is preceded by Kozak sequence GCCACCATGG.

3-8. -8. (canceled)

16. A method for treating a dust mite allergic disease, comprising administering a recombinant Der p2 protein having an amino acid sequence as shown in SEQ ID NO:

3 to a subject to be treated.