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Recombinant fusion protein (vaccine) compsn. contg. same and method for prodn. thereof

A technology of vaccine composition and fusion protein, which is applied in the field of recombinant fusion protein and can solve the problems of thorough purification and other problems

Inactive Publication Date: 2006-07-19
LOHMANN ANIMAL HEALTH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, both the preparation and thorough purification of St×2e were problematic

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0065] Preparation of St×2e recombinant B subunit

[0066] For the preparation of recombinant B subunits, the strain E. coli Cux-St×2eB, DSM number 12721 (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg Ib, D-38124 Brauschweig) was used. This laboratory strain of E. coli contains the plasmid pHIT-24 from which the B-subunit of Stx2e is cloned.

[0067] A seed batch system was established from this strain and refrigerated at -78°C in 2ml cryogenic vials.

[0068] To generate recombinant B subunits, 1 ampoule of working seed (2 ml) was thawed to grow preculture 1 .

[0069] The preparation conditions of preculture 1 were as follows:

[0070] Culture medium: 150ml of sterile standard I nutrient broth + 0.01% ampicillin was put into a 30ml Erlenmeyer flask. Cultivation: 37°C, 15 hours, upright and static culture.

[0071] The main substance was prepared using a fermentor "BIOSTAT B" with a 5 liter culture vessel. The culture vessel was filled with...

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Abstract

The invention relates to a recombinant fusion protein, containing a subgenic Stx2e fragment of the Shiga toxin 2e (Stx2e) in fusion with a terminal tag, whose size corresponds approximately to that of the fragment or to a fraction of said fragment.

Description

technical field [0001] The invention relates to a recombinant fusion protein, a (vaccine) composition containing the recombinant fusion protein and a preparation method of the recombinant fusion protein. Background technique [0002] Edema disease in pigs is caused by Shiga toxin-forming Escherichia coli (STEC). The only major virulence factor of this pathogen responsible for this clinical disease is Shiga toxin 2e (St x 2e) (MacLeod et al., 1991). Since the onset of the disease is acute in most cases and treatment is often initiated too late or does not achieve the expected results, it is of great interest to develop an effective preventive method. However, both the preparation and thorough purification of Stx2e were problematic. [0003] The B subunit of St×2e has been considered as a vaccine candidate for various reasons. It was identified from the serum of recovered piglets, ie it possessed the epitope. Second, the B-subunit of the toxin induces the formation of toxi...

Claims

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Application Information

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IPC IPC(8): C07K14/245C12N15/62A61K38/16C07K16/12C12N15/63C12N15/06A61K39/00
CPCC07K14/245A61K39/00C07K2319/00
Inventor G·巴尔杰S·弗兰克
Owner LOHMANN ANIMAL HEALTH