Pneumococcal surface proteins

A pneumococcal and protein technology, applied in antibacterial drugs, bacterial antigen components, allergic diseases, etc., can solve problems such as easy deamination

Pending Publication Date: 2021-11-09
THE UNIV OF TOKYO +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, PspA derived from the D39 strain is prone to deamination at a pH near neutral, and has a problem in terms of stability.

Method used

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  • Pneumococcal surface proteins
  • Pneumococcal surface proteins
  • Pneumococcal surface proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0062] 1. Native PAGE

[0063]Various PspA proteins, PspA 1-1 (SEQ ID NO: 5), PspA 1-2 (SEQ ID NO: 6), PspA 1-3 (SEQ ID NO: 4) and PspA 1-3LB (SEQ ID NO: 3) were expressed in Escherichia coli Medium expression (refer to WO2018102774 for details). The expressed PspA protein was extracted non-denaturingly by conventional methods, and purified using ion-exchange chromatography, butyl agarose, etc. until it became homogeneous in LDS (Lithium Dodecyl Sulfate)-PAGE.

[0064] First, PspA 1-3 molecules were suspended in 2 mg / ml PBS immediately after purification and washed with 0.5M NaH 2 PO 4 or 0.5MNa 2 HP 4 The pH was adjusted to a predetermined pH (pH6.5, pH7.0, pH7.5, pH8.0, and pH8.5), and stored at 25°C for a predetermined time (1 day, 2 days, and 5 days). 10 μl (1.0 μg / lane) of each sample was electrophoresed by Native PAGE at 200 V for 60 minutes, and stained with Coomassie blue (BioRad). It should be noted that the gel used was 7.5% Mini-Protein TGX Gel12well (BioRad),...

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Abstract

The present invention provides D39-derived mutant PspA that does not undergo deamination and maintains molecular stability even at near-neutral pH. Specifically, the present invention is the following protein (a) or (b). (a) A protein that includes the amino acid sequence represented by sequence no. 2 and that has pneumococcus vaccine antigenic activity, and a protein substantially the same as said protein; and (b) a protein that includes a portion of the amino acid sequence represented by sequence no. 2, said portion including aspartic acid at position 254, and that has pneumococcus vaccine antigenic activity, and a protein substantially the same as said protein.

Description

technical field [0001] The present invention relates to pneumococcal surface protein A (PspA) and vaccines comprising PspA. Background technique [0002] Pneumococcus is a clinically important pathogen of upper respiratory tract infection alongside influenza virus. It is complicated by otitis media to pneumonia, bacteremia, meningitis, etc., and causes serious illness including death in children and adults. [0003] In recent years, 7-, 10-, and 13-valent polysaccharide-conjugated pneumococcal vaccines (PCV (pneumococcal conjugate vaccine) 7, PCV10 and PCV13), administered by intramuscular injection. However, this polysaccharide-based vaccine hardly induces an immune response in children due to the weak immunogenicity of the T-cell-independent polysaccharide, and also shows an infection defense effect only against capsular serotype pneumococci. Furthermore, since intramuscular injection of vaccines mainly induces systemic anti-IgG antibodies, these vaccines have problems s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/09A61P11/00A61P37/04C07K14/315C12N15/31
CPCA61P11/00C07K14/315A61K39/092A61P31/04A61K2039/575A61K2039/57A61K2039/54A61K2039/545C07K14/3156
Inventor 幸义和中桥理佳清野宏
Owner THE UNIV OF TOKYO
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