The limited success of current treatment options for Glioblastoma, an aggressive brain cancer with poor survivability can be attributed to the blood-brain barrier (BBB). This barrier is known to prevent the entry of most chemotherapeutic drugs to treat brain cancer thus, a novel targeted therapeutic capable of crossing the BBB to deliver drugs is essential. Aptamers, or chemical antibodies, are small single-stranded nucleotide sequences that can bind specifically and selectively to desired targets on the cell surface. Most importantly, aptamers can be modified as a drug delivery vehicle for therapeutic purposes. In this instance, we previously generated a bifunctional aptamer by combining a transferrin receptor and epithelial cell adhesion molecule (EpCAM) aptamer together and intercalated the chemotherapeutic doxorubicin (DOX) for the treatment of brain metastases. We were able to demonstrate that the bifunctional aptamer-drug conjugate, termed TEPP-DOX, was successfully able to deliver drug payloads across the BBB to EpCAM positive brain metastases both in vitro and in vivo, and reduce metastases spread and tumourigenicity [1]. In this study, for the first time, this bifunctional aptamer-drug conjugate is being tested for treating the primary brain cancer, Glioblastoma. To examine the potential of treating Glioblastoma, the binding affinity of two bifunctional aptamers to Glioblastoma was determined by flow cytometry, including with DOX conjugation, where a strong binding affinity towards the transferrin receptor on Glioblastoma cells was observed. Next, the internalisation and drug retention of aptamer-DOX conjugates was visualised over a 96-hour period through confocal microscopy. The results of this study show that this aptamer-drug conjugate would make an ideal therapeutic candidate for future studies.Â