Background Enhancing the efficiency of process development is a key issue for the production of recombinant therapeutic protein. To maximize both cell mass and productivity with limited time and effort, an improved top-down approach was adopted in the development and optimization of fed-batch culture. Results Two-round optimization was carried out to refine the nutrients following an investigation into the effect of complete feeding medium. After the first round of optimization, cell growth was improved with the integral of viable cell concentration increased by 32.2%. After the second round of optimization, cell productivity was improved with the specific antibody production rate increased 1.5-fold, and cell longevity was also prolonged using improved feeding strategy. In the optimized fed-batch culture in a 2 L bioreactor, the maximum cell concentration was increased by 68% and final antibody titer was increased 3.9-fold compared with the fed-batch culture with simple feeding medium. Conclusion The improved top-down approach helps to reveal the interaction of different nutrients and their important roles in the improvement of cell proliferation and production. Thus, the common defects of a top-down approach can be avoided. This approach is practicable and satisfactory for efficient development of a high-yielding process. (c) 2013 Society of Chemical Industry