Freestanding, strip-shaped magnetoelastic (ME) biosensors are a class ofwireless, mass-based biosensors that are being developed for the real-timedetection of pathogenic bacteria for food safety and bio-security. The masssensitivity of these biosensors operating in longitudinal-vibration modes isknown to be largely dependent on the position of masses attached to thesensor surfaces. Hence, considering this dependence is crucial to thedetection of low-concentration target pathogens, including single pathogenicbacteria, because their local attachment may cause varying sensor responses.In a worst case scenario, the resultant sensor responses (i.e., mass-inducedresonance frequency changes of the sensor) may be too small to be detecteddespite the attachment of the target pathogenic masses. To address theissue, phage-coated ME biosensors (magnetostrictive strips (4 mm × 0.8 mm ×30 μm) coated with a phage probe specifically binding streptavidin protein)with localized masses (streptavidin-coated polystyrene beads) werefabricated, and mass-position-dependence of the sensor’s sensitivity underthe fundamental-mode vibration was experimentally measured. In addition,three-dimensional finite element (FE) modal analysis was performed using theCalculiX software to simulate the phenomena. The experimental andtheoretical results show close agreement: (1) the mass sensitivity was lowwhen the mass was positioned in the middle of the sensor’s longest dimensionand (2) a much higher mass sensitivity was, by contrast, obtained for theequivalent masses placed at both ends of the strip-shaped sensor.Furthermore, FE models were constructed for differently sized, phage-coatedME biosensors (100 – 500 μm in length with different widths and thicknesses)loaded with a single bacterial mass (2 μm × 0.4 μm × 0.4 μm, 1.05 g/cm3) at varying longitudinal positions. The mass sensitivitywas found to be approximated by a mass-position-dependent Boltzmann functionwhose amplitude is inversely proportional to the length squared, width, andthickness of the sensor.