Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Recombinant malaria vaccine

A kind of composition and technology of malaria parasite, applied in the field of recombinant malaria vaccine and preparation thereof

Inactive Publication Date: 2010-01-27
赫尔曼·比雅尔
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it only describes the purification of polypeptides fused to heterologous sequence tags such as GST, strep, or hexahistidine tags
However, in vaccines, the presence of such heterologous sequences is undesirable

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Recombinant malaria vaccine
  • Recombinant malaria vaccine
  • Recombinant malaria vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Preparation of p83 / 30 fragment

[0041] The amino acid sequence of the p83 / 30 fragment is shown in SEQ ID NO: 1 (strain F) or SEQ ID NO: 2 (strain D). Cloning the nucleic acid sequence encoding the fragment into figure 1 In the expression vector pZE23D 83 / 30 shown in. The fragment-encoding nucleic acid is operably linked to the IPTG-inducible pA1lacO1 promoter. The Escherichia coli production strain is W3110Z2 (for example, Bacteriol. Ref. 36, (1972), 525-530; Proc. Natl. Acad. Sci. USA 78 (1981), 7069-7072).

[0042] E. coli cells were pre-cultured in LB medium. The preculture was diluted 1:50 with Superbroth and grown in fermentors at 37°C. In optical density (OD 600 ) is 1.5, add doxycycline (200ng / ml). in OD 600 = 4.5, IPTG (1 mM) was added.

[0043] 2.5h after induction with IPTG or at OD 600 At =7, the cells were collected.

[0044] The collected bacteria were disrupted by continuous homogenization at a turnover rate of 10 l / h and a maximum pressure of 1...

Embodiment 2

[0060] Preparation of p38 / 42 fragment

[0061] The amino acid sequence of the p38 / 42 fragment is shown in SEQ ID NO: 3 (strain F) or SEQ ID NO: 4 (strain D).

[0062] Inclusion bodies of the p38 / 42 fragment were prepared in substantially the same manner as described in Example 1 for the p83 / 30 fragment.

Embodiment 3

[0064] Preparation of compositions comprising equimolar amounts of p83 / 30 and p38 / 42 fragments

[0065] The inclusion bodies obtained in Examples 1 and 2 were respectively dissolved by adding a dissolution buffer (6M guanidine hydrochloride, 50 mM sodium phosphate, 10 mM dithiothreitol, 1 mM EDTA, pH 8.0) to the inclusion body (IB) .

[0066] For the p83 / 30 fragment, three different ratios of IB to lysis buffer were used, namely 1 g IB + 2.5 ml buffer, 1 g IB + 4.0 ml buffer and 1 g IB + 8.0 ml buffer. For the p38 / 40 fragment, 1 g of IB was added to 2.5 ml of buffer.

[0067] Filtrates were refolded by incubation overnight at room temperature in 500 mM arginine, 50 mM sodium phosphate, 1 mM reduced L-glutathione, 0.1 mM oxidized glutathione, 1 mM EDTA pH 8.0.

[0068] The resulting protein solution was concentrated 5-fold by ultrafiltration. The buffer was exchanged with 20 mM sodium phosphate, 50 mM NaCl, 0.01% Tween 80, pH 8.0. After filtration with a 0.2 μm filter, the ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention refers to a recombinant malaria vaccine and a method for its manufacture.

Description

technical field [0001] The invention relates to a recombinant malaria vaccine and a preparation method thereof. Background technique [0002] The merozoite surface protein (MSP-1) of the malaria parasite (Plasmodium) is present on the surface of the merozoite, which is the erythrocyte-invasive form of the parasite. MSP-1 is produced as a precursor protein with a molecular weight of about 190 kDa, which is proteolytically processed into four fragments called p83, p30, p38, and p42 during merozoite maturation. The form of non-covalent attachment is retained on the surface of the parasite. Upon invasion of red blood cells, further proteolytic cleavage occurs. [0003] The MSP-1 protein consists of several highly conserved regions, a dimorphic region, which is grouped with one of the two allelic forms in the N-terminal region and two relatively small minority developmental regions (oligomorphic block) related to (Tanabe et al., J.Mol.Biol.195 (1987) 273-287; Miller et al., Mo...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K14/445A61K39/015
CPCA61K39/015C07K14/445A61P33/06A61P37/04Y02A50/30
Inventor 赫尔曼·比雅尔罗尔夫·卢茨克里斯琴·考思克里斯琴·埃普乌特·沃尔比尔
Owner 赫尔曼·比雅尔
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com