This study addresses the challenge of enhancing the strength of polyacrylonitrile (PAN) nanofiber mats in the transverse direction, which is a critical aspect for filtration applications targeting nanosized viruses and bacteria. Traditional methods of producing orientated nanofibers often result in mats with limited transverse strength. To overcome this, our research focused on developing biaxial nanofibers, which exhibit enhanced interbonding in both axial and transverse directions. Using a single-syringe electrospinning setup with a rotating drum collector, we successfully fabricated biaxially orientated PAN nanofiber mats. Scanning electron microscopy (SEM) analysis was employed to visualise and confirm the biaxial arrangement and porous structure of the mats. Mechanical testing further demonstrated that these biaxially arranged nanofibers possess comparable strength in both orientations, signifying a significant advancement in the structural integrity of PAN nanofiber mats for filtration applications.