The early detection of diseases through protease biomarker identification in blood samples can improve patient outcomes for various conditions, including pancreatic cancer. Traditional protease detection techniques, however, are complex, costly, and time-consuming, necessitating the processing of samples in equipped laboratories. Our research introduces a proof of concept for a novel, quantitative LFA designed to detect trypsin activity. This LFA employs a charge-changing fluorescent substrate which, upon cleavage by the protease, enables electrostatic capture of the cleaved product, distinguishing it from traditional LFAs that rely on antibodies for signal capture. The development of this LFA not only promises to enhance diagnostic efficiency and patient care but also holds the potential to transform early disease detection and management through accessible, affordable,  point-of-care testing, especially in resource-limited settings.