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Overexpression, Purification and Validation of Antigenic Salmonella enterica Serovar Typhi Proteins Identified from LC-MS/MS

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Abstract

In our earlier study, an immunoblot analysis using sera from febrile patients revealed that a 50-kDa band from an outer membrane protein fraction of Salmonella enterica serovar Typhi was specifically recognized only by typhoid sera and not sera from other febrile illnesses. Here, we investigated the identities of the proteins contained in the immunogenic 50-kDa band to pinpoint antigens responsible for its immunogenicity. We first used LC-MS/MS for protein identification, then used the online tool ANTIGENpro for antigenicity prediction and produced recombinant proteins of the lead antigens for validation in an enzyme-linked immunosorbent assay (ELISA). We found that proteins TolC, GlpK and SucB were specific to typhoid sera but react to antibodies differently under native and denatured conditions. This difference suggests the presence of linear and conformational epitopes on these proteins.

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Acknowledgments

EBO is grateful for the postdoctoral fellowship from USM during the course of this work. This work was supported by the USM Research University Cluster Grants 1001/PSKBP/8630011 and 1001/PSKBP/8630015.

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Authors and Affiliations

  1. Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Minden, Penang, Malaysia

    Chai Fung Chin, Boon Aun Teh, Eugene Boon Beng Ong & Theam Soon Lim

  2. Institute for Research in Molecular Medicine, Health Campus, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia

    Amy Amilda Anthony, Ismail Aziah & Asma Ismail

Authors
  1. Chai Fung Chin
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  2. Boon Aun Teh
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  3. Amy Amilda Anthony
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  4. Ismail Aziah
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  5. Asma Ismail
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  6. Eugene Boon Beng Ong
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  7. Theam Soon Lim
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Corresponding author

Correspondence to Eugene Boon Beng Ong.

Additional information

Chai Fung Chin and Boon Aun Teh contributed equally to this work.

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Chin, C.F., Teh, B.A., Anthony, A.A. et al. Overexpression, Purification and Validation of Antigenic Salmonella enterica Serovar Typhi Proteins Identified from LC-MS/MS. Appl Biochem Biotechnol 174, 1897–1906 (2014). https://doi.org/10.1007/s12010-014-1173-y

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  • DOI: https://doi.org/10.1007/s12010-014-1173-y

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