Bioproduced Proteins On Demand (Bio-POD) in hydrogels using Pichia pastoris

Shuo-FuYuan^a,1, Sierra M. Brooks^b,1, Annalee W. Nguyen^b, Trevor G. Johnston^c, Jennifer A. Maynard^c, Alshakim Nelson^c, Hal S. Alper^a,b

Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA.

Abstract

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.