This course provides participants with fundamental knowledge of cutting-edge cloning strategies essential for mammalian cell line development, specifically geared towards protein overproduction with specific quality attributes. With a focus on the industrial-relevant Chinese hamster ovary (CHO DG44) cells as the expression model, attendees will delve into crucial aspects such as the selection of expression vectors, understanding host cell genetics and physiology, and implementing effective screening strategies to maximize production yields. Additionally, the course highlights various facets of cell culture techniques in small-scale disposable wave bags and glass bioreactors.