Fed-batch Medium Modification for Secreted Single Chain Variable Fragment Against HIV-1 p17 Protein Production Using Combined Two Factorial Experiments in Escherichia coli Bioreactor

Abstract

Biopharmaceutical production programs have been established using fed-batch cell culture protocols, which can support high volumetric productivity while providing low operational complexity. The industry is concerned with developing high-titer cell culture processes to meet increasing market demands and reduce manufacturing costs. Although advancements in bacterial cell engineering have enabled the development of high-performing recombinant cell lines, developments in cell culture media and process parameter settings are required to realize the maximum production of those cells. We investigated two combined optimization methods, Packlett-Burman design and Sequential Simplex Optimization, for optimization of feed medium. The new combined optimization method decreased the time required by 40.7% compared to the experiment trial. Dry cell weight was 1.24 times higher than in the individual method. The fed-batch cultivation with an optimal feeding rate of 20 ml/h increased cell growth, total protein production and scFv anti-p17 activity by 4.43, 1.48 and 6.5 times compared to batch cultivation respectively.