[Objective] To improve the teaching quality of the course "Pharmaceutical bioassay technology" and the teaching ability of lecturers as well as to fully share high-quality teaching resources. [Methods] Starting from the significance of the course construction, the content was divided into 37 knowledge points; the teaching method, content and assessment of the course were discussed and analyzed with the case of "sterile examination method". [Results] The establishment of the online open course provides a vehicle for online learning and interaction for pharmaceutical students, and enriches the learning accesses of the course. [Conclusion] By reconstructing the teaching content, optimizing the teaching methods and reforming the assessment methods of this course, this study effectively enhances students' interest in learning and improve their learning effect, which can provide reference for relevant teaching reforms.