Abstract Approximately 35% of biopharmaceuticals are produced using Escherichia coli due to its rapid growth and scalability; however, recombinant proteins often accumulate as insoluble inclusion bodies, complicating recovery of bioactive forms. Despite this, inclusion bodies offer advantages such as easier isolation, protection against proteolysis, high expression levels, and the ability to express toxic proteins. The cysteine-rich hyaluronidase from Brachypelma vagans venom, exhibits specific activity against hyaluronan and has therapeutic potential. However, it is expressed as inclusion bodies in E. coli , resisting spontaneous refolding due to multiple disulfide bonds. The results herein indicate that oxidative chromatographic refolding assisted by immobilized chaperones, GroEL apical domain, DsbA, and DsbC fused to cellulose-binding modules, enables efficient recovery of active recombinant hyaluronidase from B. vagans . Over eight consecutive refolding batches, each processing 2.8 mg of denatured protein, a 100% refolding yield was achieved, surpassing conventional dialysis and dilution methods. Structural and functional analyses confirmed restoration of native-like conformation of the recombinant hyaluronidase expressed as inclusion bodies. This approach offers a scalable and effective platform for refolding disulfide bond-rich proteins from inclusion bodies, facilitating their production for pharmaceutical applications.
Antonio-Pérez et al. (Mon,) studied this question.
Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context: