Protein macroarray analysis options are thus far limited to image data processing of subsections of the array or universal manufacturer-provided scanner software packages . More dedicated software packages such as VisualGrid (GPC Biotech) or Aida Image Analyser (Raytest) rely either on operator-based decision making or fixed spot diameter measurements which are not suitable for semiquantitative comparative studies. Here we aim to develop an automated open source software package dedicated to semiquantitative analysis of protein macroarrays. We use MathWorks MATLAB version 2019b as the platform. The code is available for use in the supplementary information section.