Poster Presentation Australasian Society for Dermatology Research Annual Scientific Meeting 2024

Targeting disseminated disease using monoclonal antibody-functionalised CSPG4-targeting nanoparticles in squamous cell carcinoma (#80)

Jessica Sydney Ramos 1 2 , Jasmine Zhou 1 2 , Tzipi Cohen Hyams 1 , Chen Wang 1 2 , Kathryn Chen 1 3 , Kang Liang 4 5 , Paul de Souza 1 3 6 , Albert S. Mellick 1 4
  1. Ingham Institute for Applied Medical Research, Liverpool, NSW, Australia
  2. School of Biomedical Science, University of New South Wales, Kensington, NSW, Australia
  3. School of Medicine, Western Sydney University, Campbelltown, NSW, Australia
  4. Graduate School of Biomedical Engineering, University of New South Wales, Kensington, NSW, Australia
  5. School of Chemical Engineering, University of New South Wales, Kensington, NSW, Australia
  6. School of Medicine, University of Wollongong, Wollongong, NSW, Australia

In recent decades, there has been a rapid increase in the incidence of squamous cell carcinoma (SCCs). A certain at-risk group called epidermolysis Bullosa (EB), specifically recessive dystrophic epidermolysis bullosa (RDEB) is highly susceptible to metastatic SCCs due to their chronic wound blistering and is linked to early death. Therefore, early diagnosis is crucial for survival. An emerging type of nanoparticle technology called Zeolitic Imidazolate Framework (ZIF)-8 is comprised of a metallic ligands network bound to Zn2+ transition metal ions. ZIF-8s can be functionalised with enzymes and monoclonal antibodies (mAbs) which target an SCC marker called chondroitin sulphate proteoglycan 4 (CSPG4) for cell recognition and separation purposes. Despite this, the use of ZIF-8s in cell separation is a newly introduced method with limited data on capturing circulating tumour cells (CTCs). Further, commercial mAbs are a costly, rate-limiting step with questionable validity. Here, we conducted the optimisation of in-house production, purification using affinity chromatography and conjugation of 9.2.27 mAbs for surface-functionalisation. Further, we demonstrated the characterisation and optimisation of enzyme-encapsulated ZIF-8s, using glucose oxidase (GoX) and catalase (CAT), with embedded 9.2.27 mAbs. Then, we performed cell recovery in SCC spiked-in experiments in homogenous populations and analysed results using FACs analysis. As a result, purified, commercial grade labelled in-house mAbs were obtained. The characterised mAb ZIF-8 nanoparticles were rhombic shaped, with a mean length of 45nm, 61% enzyme encapsulation and mAb loading on ZIF-8 of 85.7%. For cell recovery, the Wild-type (WT) SCC cell line (WT-A431) had the highest cell recovery 84.06% whilst RDEB-62SCC observed a significantly lower recovery rate of 0.07%. The present study provides preliminary evidence that CSPG4-targeting ZIF-8 nanoparticles are a potential candidate for cell separation in SCC diagnostics.