Human chaperonin 10 (Cpn10) and chaperonin 60 (Cpn60) fulfil an essential role in mitochondrial protein folding. Cpn10 is located in the mitochondrial matrix; however, it has also been detected in extra-mitochondrial compartments where it has demonstrated anti-inflammatory and immunoregulatory activity, suggesting a potential therapeutic value for the treatment of inflammatory and autoimmune disorders that corresponds with its recently proposed role as a resolution-associated mol. patterns (RAMPs) mol. Ala-Cpn10, a recombinant, minimally modified Cpn10 synthesized in Escherichia coli, is formulated for use in clin. trials and pre-clin. studies with i.v. or s.c. administration. Herein, we report the development of a bioprocess for the production of ∼115 g of recombinant human Ala-Cpn10 from a 100 L E. coli fermentation with >99% purity (SDS-PAGE), <0.6 EU mg-1 endotoxin, <18 pg mg-1 DNA and 3.2% mol. variants. This bioprocess was achieved through a careful optimization of the gene construct, the promoter system and the fermentation process. Recombinant Ala-Cpn10 produced in this process is active as a mol. chaperone indicating correct tertiary and quaternary structure and the stability profile indicates no significant changes with storage as a liquid for at least 3 years at 2-8 °C. The results of validated characterization assays demonstrate that purified Ala-Cpn10 produced using the optimized process reported here is suitable for its intended purpose as an investigational drug product, and this material is currently being tested in a phase II study for efficacy, safety and impact on biomarkers in subjects with mildly active Systemic Lupus Erythematosus (SLE) under US IND 116156.