The purpose of this observational study was to compare anthropometric and functional characteristics of office worker women who were healthy or suffered from chronic low back pain (CLBP). A group of 30 healthy and 30 women with CLBP were randomly selected from a sample of 648 office workers who participated in a previous epidemiological study. All subjects were matched for age, height and body weight. Their anthropometric and functional characteristics were measured using standardized procedures with established reliability and compared with the Student’s t-test for independent samples. In their anthropometric characteristics, women with CLBP had significantly higher body mass index and percent body fat (p = 0.035) whereas their leg length difference (p = 0.012) was almost double compared to same characteristics of healthy women. Regarding their functional characteristics, women with CLBP displayed significantly restricted mobility (p < 0.05) in all directions and decreased endurance (p < 0.05) in muscle function of their lower extremity, abdominals and trunk extensors as compared to healthy women. These results might suggest that anthropometric and functional characteristics need to be improved in Greek office workers as they may relate to the incidence of their low back pain.