Ranibizumab is an antibody fragment used for the treatment of blurred vision due to age-related macular degeneration. Most manufacturers currently express it in Escherichia coli BL21 (DE3), where low-level expression of this complex protein has been acknowledged to result in higher production cost. This work aims to present a strategy involving the use of additives to enhance the expression of Ranibizumab in E. coli. The effect of ethanol (as additive) on cell size, cellular physiology and cellular viability were examined. A two-fold improvement in Ranibizumab expression has been demonstrated using the optimized conditions vis-a-vis the control, thereby demonstrating the efficacy of the proposed approach. LC-MS confirmed correct disulfide bond formation and Surface plasmon resonance confirmed the formation of active recombinant protein via binding to its target VEGF. Transcriptomic analysis and RT-qPCR validation indicated that changes in membrane properties and DNA synthesis results in growth, gene amplification and enhances synthesis of inducible proteins in case of the optimized medium.

Ethanol as an additive enhances expression of Ranibizumab in Escherichia coli: Impact on cellular physiology and transcriptome

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.