Production of a Recombinant Vaccine Candidate against Burkholderia pseudomallei Exploiting the Bacterial N-Glycosylation Machinery

Frontiers in Microbiology

卷:5

Chad eStratilo¹  Jeremy Andrew Iwashkiw²  Nancy L Price²  Mario eFeldman²  Fatima eGarcia-Quintanilla² 

[1] Defence Research and Development Canada Suffield;

[2] University of Alberta;

关键词: Mass Spectrometry;    Molecular Biology;    Vaccines;    Glycobiology;    protein glycosylation;    Microbiology and Biotechnology;   

DOI  :  10.3389/fmicb.2014.00381

来源: DOAJ

【 摘 要 】

Vaccines developing immune responses towards surface carbohydrates conjugated to proteins are effective in preventing infection and death by bacterial pathogens. Traditional production of these vaccines utilizes complex synthetic chemistry to acquire and conjugate the glycan to a protein. However, glycoproteins produced by bacterial protein glycosylation systems are significantly easier to produce, and could possible be used as vaccine candidates. In this work we functionally expressed the B. pseudomallei O polysaccharide (OPS II), the C. jejuni oligosaccharyltransferase (OTase), and a suitable glycoprotein (AcrA) in a designer E. coli strain with a higher efficiency for production of glycoconjugates.We were able to produce and purify the OPS II-AcrA glycoconjugate, and MS analysis confirmed correct glycan was produced and attached.We observed the attachment of the O-acetylated deoxyhexose directly to the acceptor protein, which expands the range of substrates utilized by the OTase PglB.Injection of the glycoprotein into mice generated an IgG immune response against B. pseudomallei, and this response was partially protective against an intranasal challenge.Our experiments show that bacterial engineered glycoconjugates can be utilized as vaccine candidates against B. pseudomallei. Additionally, our new E. coli strain SDB1 is more efficient in glycoprotein production, and could have additional applications in the future.

【 授权许可】

Unknown