Source:http://linkedlifedata.com/resource/pubmed/id/20937316
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
51
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| pubmed:dateCreated |
2010-12-3
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| pubmed:abstractText |
Polysaccharide-based vaccines against Neisseria meningitidis (Nm) serogroups A, C, Y and W135 have been available since 1970, but similar vaccine candidates developed for Nm group B (NmB) have not been successful due to both poor immunogenicity and their potential immunological cross-reactivity with human neurological tissue. In previous reports, a protective antigen and vaccine candidate, Ag473, was identified using proteomics and NmB-specific bactericidal monoclonal antibody. To initiate human phase one clinical trials, antigen production and characterization, pre-clinical toxicology and animal studies are required. In the present study, we report the biochemical characterization of Escherichia coli-expressed recombinant Ag473 (rAg473). Using MALDI-TOF mass analysis, chromatographically purified rAg473 was found to have two major isoforms that have molecular masses of 11,306 and 11,544amu, respectively. The isoforms were separated using RP-HPLC and pooled into two fractions. Based on the chromatogram, the ratio of lipoproteins in fractions #1 and #2 was found to be 1-2. GC-MS analysis of lipoproteins was performed, and the acylated fatty acids were identified. The results indicated that the first lipoproteins in fraction #1 contained the lipids palmitic acid (C16:0), cyclopropaneoctanoic acid (C17:1) and, predominately, stearic acid (C18:0). A different lipid composition of cyclopropaneoctanoic acid (C17:1), oleic acid (C18:1) and, predominately, palmitic acid (C16:0) was found in the second lipoprotein fraction. Both lipoprotein isoforms were tested and found to have Toll-like receptor (TLR) agonist activity in stimulating cytokine secretion from THP-1 cells. Circular dichroism (CD) analysis showed the secondary structure of rAg473 to be dominated by ?-helices (48%), and the overall protein structure was stable up to 60°C and could refold after having been exposed to a temperature cycle from 20 to 90°C. In addition, the solubility of rAg473 (5mg/mL) was not affected after several freeze-thaw cycles. These biophysical and immunological properties make rAg473 a good vaccine candidate against NmB.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies, Bacterial,
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, Bacterial,
http://linkedlifedata.com/resource/pubmed/chemical/Bacterial Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Fatty Acids,
http://linkedlifedata.com/resource/pubmed/chemical/Immunoglobulin G,
http://linkedlifedata.com/resource/pubmed/chemical/Lipoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/NMB1468 protein, Neisseria...,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Isoforms,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Toll-Like Receptors
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| pubmed:status |
MEDLINE
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| pubmed:month |
Nov
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| pubmed:issn |
1873-2518
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| pubmed:author |
pubmed-author:ChongPeleP,
pubmed-author:ChouAi-HsiangAH,
pubmed-author:HsiehShih-YangSY,
pubmed-author:KwokYanY,
pubmed-author:LengChih-HsiangCH,
pubmed-author:LinChang-LingCL,
pubmed-author:LinLi-HsiuLH,
pubmed-author:LiuShih-JenSJ,
pubmed-author:MILTONT HTH,
pubmed-author:SungJerry Wang-ChouJW,
pubmed-author:YangChiou-YingCY
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| pubmed:copyrightInfo |
Copyright © 2010 Elsevier Ltd. All rights reserved.
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| pubmed:issnType |
Electronic
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| pubmed:day |
29
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| pubmed:volume |
28
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
8175-82
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| pubmed:meshHeading |
pubmed-meshheading:20937316-Animals,
pubmed-meshheading:20937316-Antibodies, Bacterial,
pubmed-meshheading:20937316-Antigens, Bacterial,
pubmed-meshheading:20937316-Bacterial Proteins,
pubmed-meshheading:20937316-Chromatography,
pubmed-meshheading:20937316-Chromatography, High Pressure Liquid,
pubmed-meshheading:20937316-Circular Dichroism,
pubmed-meshheading:20937316-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:20937316-Escherichia coli,
pubmed-meshheading:20937316-Fatty Acids,
pubmed-meshheading:20937316-Gas Chromatography-Mass Spectrometry,
pubmed-meshheading:20937316-Gene Expression,
pubmed-meshheading:20937316-Hot Temperature,
pubmed-meshheading:20937316-Immunoglobulin G,
pubmed-meshheading:20937316-Lipoproteins,
pubmed-meshheading:20937316-Mice,
pubmed-meshheading:20937316-Mice, Inbred BALB C,
pubmed-meshheading:20937316-Molecular Weight,
pubmed-meshheading:20937316-Neisseria meningitidis, Serogroup B,
pubmed-meshheading:20937316-Protein Conformation,
pubmed-meshheading:20937316-Protein Isoforms,
pubmed-meshheading:20937316-Protein Stability,
pubmed-meshheading:20937316-Recombinant Proteins,
pubmed-meshheading:20937316-Spectrometry, Mass, Matrix-Assisted Laser...,
pubmed-meshheading:20937316-Toll-Like Receptors
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| pubmed:year |
2010
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| pubmed:articleTitle |
Biochemical characterizations of Escherichia coli-expressed protective antigen Ag473 of Neisseria meningitides group B.
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| pubmed:affiliation |
Vaccine Research and Development Center, National Health Research Institutes, Zhunan Town, Miaoli County 350, Taiwan.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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