The aim of this study was to optimise the 64Cu‐labelling of DOTA‐trastuzumab and as the first to produce and compare with its 1,4,7‐triazacyclononane, 1‐glutaric acid‐5,7 acetic acid (NODAGA) analogue in a preliminary HER2 tumour mouse model. Here, ERBB2 is linked to neoplasm.