Nowadays, medical rehabilitation system has become a requirement due to increment in national rehabilitation centres and medical hospitals. An assistive rehabilitation orthosis becomes essential and was used for rehabilitation therapy, condition monitoring, and physical strengthening. This study focused on the lower limb assistive rehabilitation orthosis development using pneumatic artificial muscle. To successfully control this orthosis system which consists of antagonistic mono-and biarticular muscle actuators, it is necessary to construct a reliable control algorithm. The suitable control scheme and strategy to manoeuvre this orthosis system similar to human musculoskeletal system have yet to be fully developed and established. Based on the review study, it is said that the co-contraction controls of anterior-posterior pneumatic muscles was able to improve the joint stiffness and stability of the orthosis as well as good manoeuvrability. Therefore, a characterization model of an antagonistic mono-and bi-articular muscles activities of human's lowerlimb during walking motion will be necessary. A healthy young male subject was used as test subject to obtain the sEMG muscle activities for antagonistic mono-and bi-articular muscles (i.e., Vastus Medialis-VM, Vastus Lateralis-VL, Rectus Femoris-RF, and Bicep Femoris-BF). The tests were carried out at different speeds of 2km/h, 3km/h, and 4km/h for one minute walking motion on a treadmill. Then, the patterns of the sEMG muscle activities were modelled and characterised using fifth order polynomial equation. Based on the results, it is shown that the anterior and posterior muscles were exhibited a muscle synergy in-between multiple anterior or posterior muscles and muscle co-contraction between anteriorposterior muscles in order to control the movements at the joints during walking motion. As conclusion, it is proven that the sEMG muscle activities of the antagonistic mono-and bi-articular muscles were follow a certain contraction-expansion patterns during walking motion even when it were tested at different gait cycle speeds.