Development Of Tcr-like Single Domain Antibody By Incorporating Immunoglobulin Isotypes For Latent Tuberculosis Diagnostics And Therapeutics Application Against Heat Shock Protein 16 Kda Peptide Presented By Hla-a2
Heat shock protein 16 kDa (HSP 16 kDa) is essential for the survival of latent Mycobacterium tuberculosis. Therefore, a peptide-MHC presentation of HSP 16 kDa could be a potential diagnostic and therapeutic target for latent tuberculosis (LTB). This study aimed to generate different immunoglobuli...
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| Format: | Thesis |
| Language: | English |
| Published: |
2024
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| Subjects: | |
| Online Access: | http://eprints.usm.my/62136/ http://eprints.usm.my/62136/1/LIU%20HUAQIANG%20-%20TESIS24.pdf |
| Summary: | Heat shock protein 16 kDa (HSP 16 kDa) is essential for the survival of latent
Mycobacterium tuberculosis. Therefore, a peptide-MHC presentation of HSP 16
kDa could be a potential diagnostic and therapeutic target for latent tuberculosis
(LTB). This study aimed to generate different immunoglobulin isotype pairings on a
TCR-like single-domain antibody (sDAb) and subsequently investigate its
diagnostic and therapeutic potential in LTB, utilizing a model cell presenting the
target peptide. A previously generated TCR-like sDAb that can bind to HSP 16 kDa
was first fused to a human IgG1, IgG2, IgG4, IgA, IgD, IgE, and IgM Fc-receptor
via a linker. The fusion product, sDAb-Fc, was expressed in HEK293-F cells and
subsequently purified. Its diagnostic potential was investigated via cell-based
ELISA utilizing MCF-7 cells peptide-pulsed with HSP 16 kDa peptides.
Antibody-dependent cell-mediated cytotoxicity (ADCC) of MCF-7 cells was also
conducted to investigate its therapeutic potential. Finally, TCR-like sDAb-IgG1,
IgG2, and IgG4 were successfully produced transiently in HEK293 F cells and
purified using protein A chromatography. |
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