Identification of antigenic epitopes of S. typhi using phage display epitopes library

We have constructed a genome-targeted library of Salmonella typhi displayed on the surface of filamentous phage particles. The genome-targeted library was made by cloning and expressing relatively short DNA fragments (100-300bp) from S. typhi genomic DNA in the pIII phage coat protein genes of a pha...

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Main Authors: Thong, Kwai Lin, Subramaniam, G., Devi, S., Puthucheary, S.D., Yu, M., Wang, L.F., Pang, T.
Format: Article
Language:English
Published: 1998
Subjects:
Online Access:http://eprints.um.edu.my/10011/1/Thong_et_al.%2C_1998.pdf
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author Thong, Kwai Lin
Subramaniam, G.
Devi, S.
Puthucheary, S.D.
Yu, M.
Wang, L.F.
Pang, T.
author_facet Thong, Kwai Lin
Subramaniam, G.
Devi, S.
Puthucheary, S.D.
Yu, M.
Wang, L.F.
Pang, T.
author_sort Thong, Kwai Lin
collection UM
description We have constructed a genome-targeted library of Salmonella typhi displayed on the surface of filamentous phage particles. The genome-targeted library was made by cloning and expressing relatively short DNA fragments (100-300bp) from S. typhi genomic DNA in the pIII phage coat protein genes of a phagemid. Utilizing a biopanning assay, where diluted sera from patients with typhoid fever were immobilized on a solid support (paramagnetic beads), antigenic epitope from the genome-targeted phage library were identified following binding and subsequent elution of recombinant phages and DNA sequencing of relevant inserts. Database searching of the identified sequence was carried out and putative antigenic epitopes identified. The power of this approach lies in its ability to search for entire spectrum of antigenic epitopes and in assessing individual patient's immune responses to particular strains of S. typhi. The findings may have important implications for improved understanding of disease pathogenesis, better diagnostic and future development of vaccines.
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spelling um.eprints-100112018-10-12T06:31:28Z http://eprints.um.edu.my/10011/ Identification of antigenic epitopes of S. typhi using phage display epitopes library Thong, Kwai Lin Subramaniam, G. Devi, S. Puthucheary, S.D. Yu, M. Wang, L.F. Pang, T. QR Microbiology We have constructed a genome-targeted library of Salmonella typhi displayed on the surface of filamentous phage particles. The genome-targeted library was made by cloning and expressing relatively short DNA fragments (100-300bp) from S. typhi genomic DNA in the pIII phage coat protein genes of a phagemid. Utilizing a biopanning assay, where diluted sera from patients with typhoid fever were immobilized on a solid support (paramagnetic beads), antigenic epitope from the genome-targeted phage library were identified following binding and subsequent elution of recombinant phages and DNA sequencing of relevant inserts. Database searching of the identified sequence was carried out and putative antigenic epitopes identified. The power of this approach lies in its ability to search for entire spectrum of antigenic epitopes and in assessing individual patient's immune responses to particular strains of S. typhi. The findings may have important implications for improved understanding of disease pathogenesis, better diagnostic and future development of vaccines. 1998 Article PeerReviewed application/pdf en http://eprints.um.edu.my/10011/1/Thong_et_al.%2C_1998.pdf Thong, Kwai Lin and Subramaniam, G. and Devi, S. and Puthucheary, S.D. and Yu, M. and Wang, L.F. and Pang, T. (1998) Identification of antigenic epitopes of S. typhi using phage display epitopes library. Medical Journal of Indonesia, 7. pp. 181-183.
spellingShingle QR Microbiology
Thong, Kwai Lin
Subramaniam, G.
Devi, S.
Puthucheary, S.D.
Yu, M.
Wang, L.F.
Pang, T.
Identification of antigenic epitopes of S. typhi using phage display epitopes library
title Identification of antigenic epitopes of S. typhi using phage display epitopes library
title_full Identification of antigenic epitopes of S. typhi using phage display epitopes library
title_fullStr Identification of antigenic epitopes of S. typhi using phage display epitopes library
title_full_unstemmed Identification of antigenic epitopes of S. typhi using phage display epitopes library
title_short Identification of antigenic epitopes of S. typhi using phage display epitopes library
title_sort identification of antigenic epitopes of s typhi using phage display epitopes library
topic QR Microbiology
url http://eprints.um.edu.my/10011/1/Thong_et_al.%2C_1998.pdf
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