The efficiency of the protein refolding process lies in identification of the optimal conditions. However, a number of challenges need to be overcome to achieve this. This review first describes the protein refolding process that is utilized presently for production of protein therapeutics. Next, it discusses the various shortcomings that exist with respect to the present approach. The focus of the paper is on presentation of the significant advancements that have been made in the past decade in the various aspects of protein folding, including use of bioinformatics, mechanistic modeling, analytical monitoring, process optimization, use of additives, high throughput development, on-column refolding, Quality by Design (QbD), Process Analytical Technology (PAT), and process intensification. Finally, an approach is proposed that incorporates the best practices that have been identified in the various areas. The paper is expected to be of interest to those in academia and industry working in the area of protein refolding. (C) 2013 Society of Chemical Industry