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: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2024
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| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/46408/3/Thesis%20Ms._Nur%20Izzah%20bt%20Ismadol.pdf |
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| Summary: | 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. |
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