Abstract Host-plant resistance (HPR) remains a vital tool to manage soybean aphid (Aphis glycines Matsumura), a major pest of soybean in Midwestern United States and southern Canada. HPR can be overcome by virulent biotypes of A. glycines; thus, in order to increase the durability of resistant cultivars, HPR needs to be deployed strategically. To improve the strategic deployment, a complete understanding of HPR in existing resistant germplasm will help ensure HPR success. In this study, we characterized HPR soybean to determine antibiosis and antixenosis categories of resistance to different biotypes of A. glycines. No-choice and free-choice tests were performed on 11 previously reported plant introductions (PIs) possessing resistance to at least one A. glycines biotype (1, 2, and 3). Overall, we found that the PIs manifested differences of a particular resistance category in response to infestation by different biotypes. Our data from no-choice tests indicate that all tested PIs possess antibiosis-based resistance to three biotypes. However, the strength of antibiosis was variable as some PIs showed stronger antibiosis toward a given biotype than others. All tested PIs manifested antixenosis, in addition to antibiosis. Furthermore, detached leaf assays revealed that resistance to A. glycines was not retained in excised soybean leaves. Characterization of resistance in this study can contribute to develop strategies for future deployment of resistant cultivars developed from these PIs.