The aim of this study was to examine gender differences in match performance characteristics of elite soccer players. Fifty-four male and fifty-nine female soccer players were tracked during UEFA Champions League matches using a multi-camera system (Amisco, Nice, France). Male players covered more (P < .01) distance than female players in total during a match (Effect Size [ES]: 0.5) and at higher speed thresholds (>15, >18, 18–21, 21–23, 23–25 and >27 km h−1; ES: 0.7–1.4). Decrements in the second versus first half (P < .01) were only evident in female players for the distance covered in total and at selected speed thresholds (12–15, >12 and >15 km h−1; ES: 0.6). Male central midfielders covered more (P < .01) total distance during a match than female central midfielders and at selected speed thresholds (15–23 km h−1; ES: 1.3–2.2). Male full-backs and wide midfielders covered a greater distance (P < .01) than female players in the same positions at higher speed thresholds (>15, 21–23, 23–25, 25–27 and >27 km h−1; ES: 1.5–3.1). The distance covered during the most intense 5 min period of the match (>15 km h−1) was higher (P < .01) in male compared to female players (ES: 1.0) but no distance deficit in the next versus the average 5 min period was observed for either gender (ES: 0.1–0.2). No gender differences were found for technical events such as the number of ball touches, time in possession of the ball or total duels won during both halves and the entire match (ES: 0.1–0.3). However, female players lost the ball more often (P < .05) and displayed lower pass completion rates than male players during both halves and the entire match (ES: 0.5–0.9). The data demonstrate that large gender differences exist for match performance characteristics of players competing at the highest competitive standard of European soccer. Such detailed analyses could be useful for gender-specific training information for optimal preparation. However, more research is warranted to establish gender-specific speed thresholds for elite soccer players.