Bioproduced Proteins On Demand (Bio-POD) in hydrogels using Pichia pastoris
Shuo-FuYuana,1, Sierra M. Brooksb,1, Annalee W. Nguyenb, Trevor G. Johnstonc, Jennifer A. Maynardc, Alshakim Nelsonc, Hal S. Alpera,b
Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA.
Traditional production of industrial and therapeutic proteins by eukaryotic cells typically requires large-scale fermentation capacity. As a result, these systems are not easily portable or reusable for on-demand protein production applications. In this study, we employ Bioproduced Proteins On Demand (Bio-POD), a F127-bisurethane methacrylate hydrogel-based technique that immobilizes engineered Pichia pastoris for preservable, on-demand production and secretion of medium- and high-molecular weight proteins (in this case, SEAP, α-amylase, and anti-HER2). The gel samples containing encapsulated-yeast demonstrated sustained protein production and exhibited productivity immediately after lyophilization and rehydration. The hydrogel platform described here is the first hydrogel immobilization using a P. pastoris system to produce recombinant proteins of this breadth. These results highlight the potential of this formulation to establish a cost-effective bioprocessing strategy for on-demand protein production.
Keywords: Protein production, Hydrogel, Pichia pastoris, Immobilization, Lyophilization.