| Summary: | Antibody Drug Conjugates (ADC), are biopharmaceutical drugs that are used in cancer treatment. It comprises of an antibody portion that targets the cancer cells and a conjugated cytotoxic drug to kill them. This project aims to develop a novel approach towards ADC production by utilizing Butelase1 (a peptide ligase) to specifically modify Herceptin enzymatically. Traditional methods of manufacturing ADC involves chemical modifications which introduces heterogeneous products of varying antibody-to-drug ratio, that may be toxic. In this FYP, attempts to express His-tag-TEV-Her2-GS-NHV (fusion protein of Herceptin heavy chain with NHV recognition motif and his-tag) was made via molecular cloning, lentiviral packaging, mammalian cell culture and protein purification. Results showed that the molecular cloning to produce recombinant plasmid carrying the His-tag-TEV-Her2-GS-NHV gene was successful. However, the expression of His-tag-TEV-Her2-GS-NHV protein was inconclusive as it could not be detected via western blotting. Moreover, Butelase1-mediated modifications experiments conducted were not successful indicating that there could have been complications with regards to purification or expression of His-tag-TEV-Her2-GS-NHV protein. In spite of that, this FYP serves as a positive step forward to spearhead efforts in developing an exquisite approach for homogeneous ADC manufacturing.
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