We are pleased to present two students who are close to the end of their PhD here at AIBN to share their research with you.
Date: Thursday, 21 Aril
Time: 12.30 - 1.30pm
Venue: Online Via Zoom
Click here to access the free seminar.
Development of Bispecific Antibody Deliveries Against Intracellular Targets
Ms Suchada Niamsuphap, Australian Institute for Bioengineering and Nanotechnology, UQ.
Approved therapeutic antibodies currently only target cell surface molecules or extracellular proteins. Despite high antigen specificity and affinity, the use of antibodies in targeting intracellular proteins is limited. This is because of their inability to cross the plasma membrane, and their susceptibility to proteolytic degradation by endolysosomes. Development of strategies that enable antibodies to internalise into cells and escape from endosomes would expand therapeutic targets, particularly in oncological settings.
Here, we used a protein-based drug delivery system, bispecific antibodies (bsAbs), which are tandem single-chain Fv (scFv) conjugated to endosomolytic peptides. Engineered bsAbs are a combination of a cell receptor targeting binding scFv and intracellular therapeutic scFv. The former scFv targets epidermal growth factor receptors that enable cytosolic delivery via receptor-mediated endocytosis. Once inside the cell, the latter therapeutic scFv interacts with its antigen — the SOX18 transcription factor — to block subsequent gene activation and cell proliferation. The combination of therapeutic and cell surface targeting moieties not only ensures tumour selectivity and cellular uptake.
The use of endosomolytic peptides to enhance intracellular translocation and endosomal escape efficiency of bsAbs were evaluated via luciferase-based reporter assay. In addition, the measurement of binding affinity and kinetic analysis of antibody-antigen interactions against EGFR and SOX18 proteins were assessed. Flow cytometry and confocal laser scanning microscopy analysis were also employed to ensure that the antibodies can bind to cancer cells via EGFR and internalise into cells, respectively. Our findings may lead to an early prototype of an effective cytosolic delivery and release system.
About Suchada:
Mai (Suchada) is a third year PhD student working on antibody deliveries for intracellular targets. She is a pharmacist with a keen interest in biopharmaceutical R&D and manufacturing. She graduated with Doctor of Pharmacy (PharmD) from Chulalongkorn University, where she majored in Pharmaceutics and Industrial Pharmacy. For her undergraduate project, she formulated and developed nanoparticles for transmucosal delivery. Suchada went on to do a Master of Biotechnology at the University of Queensland. Her final master’s project was glycoproteomic analysis of sparkling wine by LC/MS-MS at5 SCMB.
A Simple interfacial biosensing methodology to analyse DNA methylation block-copolymer remodelling and cancer progression
Mr Nicolas Constantin, Australian Institute for Bioengineering and Nanotechnology, UQ.
Screening for DNA methylation-based biomarkers has attracted increasing attention because of their early occurrence during cancer, their presence in most body fluids, and their ability to provide clinically relevant information for cancer diagnostic and treatment monitoring. We recently proposed an innovative methodology to detect Methylscape – a newly discovered cancer biomarker characterized by a specific DNA methylation landscape across the genome. This global epigenetic signature unique to most cancer type affects the physicochemical properties of DNA and significantly increase its gold physisorption. Based on that phenomenon we developed a straightforward interfacial biosensing approach (i.e., label-free and functionalization-free biosensor, no DNA treatment nor amplification) to electrochemically quantify DNA adsorption level at a bare gold electrode surface to differentiate healthy from cancer epigenomes. However, the previous method used a conventional three-electrode system which has limitations such as operator variability, less sensitivity and moderate reproducibility. Herein, we have optimised the methodology on a disposable Screen-Printed Electrode (SPE) system and fully characterized its biosensing precision, accuracy and reproducibility using breast cancer model cell line gDNA. The clinical utility of the developed platform to detect global epigenetic remodelling associated with cancer progression has been evaluated by profiling gDNA purified from breast and lung cancer tumour samples across different cancer stages (i.e., stage I - IV) and compared to DNA isolated from healthy controls. analysis were also employed to ensure that the antibodies can bind to cancer cells via EGFR and internalise into cells, respectively. Our findings may lead to an early prototype of an effective cytosolic delivery and release system.
About Nicolas:
Nicolas has started his PhD at AIBN in 2019 in the Centre for Personalised Nanomedicine under the supervision of Prof. Matt Trau. Before moving to Australia, he graduated from the Swiss Federal Institute of Technology in Lausanne (EPFL) with a bachelor’s degree in Life Sciences and Technologies followed by a master’s degree in Bioengineering. He experienced working in the biotech industry for a Swiss company specialized in the development of in vitro diagnostic rapid testing based on nanofluidic technology. His current research mainly focuses on electrochemical biosensing applications for the analysis of protein post-translational modifications (e.g., phosphorylation) and epigenetic biomarkers (e.g., DNA methylation) for early cancer detection and treatment monitoring.
About AIBN Seminar Series
The AIBN Seminar series showcases a range of seminars across different topics and disciplines