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Interaction of Phage Display Peptides to Chikungunya and Zika Virus Recombinant Proteins

Diagnosis by molecular methods may not be sensitive after viremic period while diagnosis by serological assay cross reactivity of antibodies might happen. Besides, serological assay usually requires specific monoclonal antibody which involves the use of animal thus raises animal welfare issues. Ther...

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Main Author: Nur Izzah, Ismadol
Format: Thesis
Language:English
Published: Society for Indonesian Biodiversity 2024
Subjects:
QR355 Virology
Online Access:http://ir.unimas.my/id/eprint/46408/3/Thesis%20Ms._Nur%20Izzah%20bt%20Ismadol.pdf
http://ir.unimas.my/id/eprint/46408/
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spelling my.unimas.ir-464082024-10-21T02:01:40Z http://ir.unimas.my/id/eprint/46408/ Interaction of Phage Display Peptides to Chikungunya and Zika Virus Recombinant Proteins Nur Izzah, Ismadol QR355 Virology Diagnosis by molecular methods may not be sensitive after viremic period while diagnosis by serological assay cross reactivity of antibodies might happen. Besides, serological assay usually requires specific monoclonal antibody which involves the use of animal thus raises animal welfare issues. Therefore, to overcome the traditional method in generating hybridoma for monoclonal antibody production, this study proposes to interrogate the phage display technology as an alternative to hybridoma screening technology for monoclonal antibody production. In this study, a Ph.D-7 Phage Display Peptide Library was used to screen high affinity ligands that could selectively bind to the purified recombinant E2 protein of CHIKV and the EDIII protein of the ZIKV. The viral antigens were immobilized to immunotubes and the phage-display library was added to the tubes to allow binding of peptide to the target molecules. The bound phage was amplified in a biopanning process. Following three rounds of biopanning, the amounts of bound phages were determined by titration. The phage clones that were positive against the viral protein in a plaque lift assay were selected and amplified. A total of 12 and 26 phage clones were amplified for potential binding with the recombinant CHIKV and ZIKV proteins respectively. The sequence analysis of the positive clones showed that, the selection with CHIKV recombinant protein generate a potential motif, NAALGT, in which 6 out of the 12 amplified clones contained the repetitive motif. However, the selection with ZIKV protein, did not show any repetitive motif sequence, the interacting peptide was further analyzed using CLUSTAL MSA software by MUSCLE suggests that these to be target unrelated peptides. Society for Indonesian Biodiversity 2024-09-27 Thesis NonPeerReviewed text en http://ir.unimas.my/id/eprint/46408/3/Thesis%20Ms._Nur%20Izzah%20bt%20Ismadol.pdf Nur Izzah, Ismadol (2024) Interaction of Phage Display Peptides to Chikungunya and Zika Virus Recombinant Proteins. Masters thesis, University Malaysia Sarawak (UNIMAS).
institution Universiti Malaysia Sarawak
building Centre for Academic Information Services (CAIS)
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Sarawak
content_source UNIMAS Institutional Repository
url_provider http://ir.unimas.my/
language English
topic QR355 Virology
spellingShingle QR355 Virology
Nur Izzah, Ismadol
Interaction of Phage Display Peptides to Chikungunya and Zika Virus Recombinant Proteins
description Diagnosis by molecular methods may not be sensitive after viremic period while diagnosis by serological assay cross reactivity of antibodies might happen. Besides, serological assay usually requires specific monoclonal antibody which involves the use of animal thus raises animal welfare issues. Therefore, to overcome the traditional method in generating hybridoma for monoclonal antibody production, this study proposes to interrogate the phage display technology as an alternative to hybridoma screening technology for monoclonal antibody production. In this study, a Ph.D-7 Phage Display Peptide Library was used to screen high affinity ligands that could selectively bind to the purified recombinant E2 protein of CHIKV and the EDIII protein of the ZIKV. The viral antigens were immobilized to immunotubes and the phage-display library was added to the tubes to allow binding of peptide to the target molecules. The bound phage was amplified in a biopanning process. Following three rounds of biopanning, the amounts of bound phages were determined by titration. The phage clones that were positive against the viral protein in a plaque lift assay were selected and amplified. A total of 12 and 26 phage clones were amplified for potential binding with the recombinant CHIKV and ZIKV proteins respectively. The sequence analysis of the positive clones showed that, the selection with CHIKV recombinant protein generate a potential motif, NAALGT, in which 6 out of the 12 amplified clones contained the repetitive motif. However, the selection with ZIKV protein, did not show any repetitive motif sequence, the interacting peptide was further analyzed using CLUSTAL MSA software by MUSCLE suggests that these to be target unrelated peptides.
format Thesis
author Nur Izzah, Ismadol
author_facet Nur Izzah, Ismadol
author_sort Nur Izzah, Ismadol
title Interaction of Phage Display Peptides to Chikungunya and Zika Virus Recombinant Proteins
title_short Interaction of Phage Display Peptides to Chikungunya and Zika Virus Recombinant Proteins
title_full Interaction of Phage Display Peptides to Chikungunya and Zika Virus Recombinant Proteins
title_fullStr Interaction of Phage Display Peptides to Chikungunya and Zika Virus Recombinant Proteins
title_full_unstemmed Interaction of Phage Display Peptides to Chikungunya and Zika Virus Recombinant Proteins
title_sort interaction of phage display peptides to chikungunya and zika virus recombinant proteins
publisher Society for Indonesian Biodiversity
publishDate 2024
url http://ir.unimas.my/id/eprint/46408/3/Thesis%20Ms._Nur%20Izzah%20bt%20Ismadol.pdf
http://ir.unimas.my/id/eprint/46408/
_version_ 1814942167519461376
score 13.222552

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