In general, larger capture areas -represented by increasing numbers of test linesresulted in notable sensitivity enhancement within an AmpliFold approach. When compared to a 'traditional' LFA example (i.e. a single, uncleaved test line), no significant difference in LOD was determined when detecting HER2 antigen using AmpliFold assays featuring single (1×) PSA test line capture strips. In this scenario, the amount of initial analyte captured is equivalent in both LFA formats, though some additional sensitivity loss is incurred by the 'capture-and-release' approach due to a small percentage of LFA format against the 'capture-and-release' AmpliFold approach under model capture affinity conditions (0.1 mg/mL PSA, low capture receptor test lines). Capture strips were printed using a varying number of test lines to probe the effects of larger capture areas on signal amplification. Data was collected in triplicate (except blanks where N=6) and plotted using a four-parameter logistic fit. d. A table collating the limit of detection for AmpliFold assays featuring different sized capture areas and a 'traditional' LFA format.