Drosophila suzukii Matsumura (Diptera: Drosophilidae) utilizes ‘Himalaya’ blackberry, Rubus armeniacus Focke (Rosaceae), as a host and may invade berry and stone fruit crops from field margins containing this invasive weed. Laboratory and semi-field studies were conducted to determine (1) the persistence of protein marks including 10% chicken egg whites (egg albumin protein), 20% bovine milk (milk casein protein), and 20% soy milk (soy trypsin inhibitor protein) on topically sprayed D. suzukii, (2) protein retention on blackberry leaves, and (3) D. suzukii acquisition of protein after exposure to marked blackberry leaves for up to 14 days after application. All flies and leaves were assayed for the presence of the protein marks using protein-specific enzyme-linked immunosorbent assays. Egg albumin, milk casein, and soy trypsin proteins persisted on 94, 49, and 25% of the topically marked D. suzukii, respectively, throughout the 14-day study period. Egg albumin was retained on 100% of treated leaves for 14 days, regardless of environmental conditions. At least 50% of flies exposed residually to egg albumin-treated leaves were marked for 3 days, regardless of exposure time and environmental conditions. However, increasing fly exposure time to treated leaves in April and June appeared to improve protein mark acquisition. Acquisition of protein by flies from treated leaves for milk casein was inconsistent, and poor for soy trypsin, despite detectable levels on treated leaves. Egg albumin had the longest and most consistent persistence on flies, leaves, and flies exposed to leaves in laboratory and semi-field studies, under a variety of environmental conditions and exposure times.