(191a) Optogenetic Platform for the “on Demand” Production of Proteins in Biopharma and Biomedical Applications

Authors: 
González-González, E., Tecnológico de Monterrey
Trujillo-de Santiago, G., Centro de Biotecnología-FEMSA, Tecnológico de Monterrey, Escuela de Ingeniería y Ciencias
Alvarez, M. M., Centro de Biotecnología-FEMSA, Tecnológico de Monterrey, Escuela de Ingeniería y Ciencias
Optogenetics, the use of light to trigger and control genetic expression, is a biological technique that is finding diverse application niches, and multiple versions of optogenetic control have been documented in recent literature.

Here, we describe a simple optogenetic platform for controlling protein expression in mammalian cell lines. Our platform uses two distinct vectors, GAV and Z-NEP. Briefly, GAV encodes for the constitutive expression of green fluorescent protein (GFP) and a fusion protein that contains a protein monomer. This last protein forms dimmers upon exposure to blue light, and dimers regulate the expression of the Z-NEP vector that encodes for a reporter (i.e., red fluorescent protein (RFP)) and/or other protein of interest (i.e., a monoclonal antibody).

The platform was tested in the CHO-S cells (the battle horse in the production of glycosylated recombinant proteins) and DU145 cells (a prostatic cancer cell line). The optogenetic control of the production of a monoclonal antibody of therapeutic interest (a biosimilar of infliximab) was demonstrated in CHO-S cells cultured in bottles and microfluidic devices. To our knowledge, this is the first report of application of optogenetics for the “on-demand” production of recombinant proteins of biopharmaceutical interest. In addition, we successfully triggered and controlled the expression of fluorescent proteins in DU145 cells exposed to blue light. We envision the control of protein expression in co-cultures of healthy and cancerous cells as a useful research and potentially therapeutic tool (i.e., controlled release mediated through light).

Our results suggest that the application of optogenetic tools to biotechnology and biomedical scenarios will enable a tight control of protein expression in organ-on-chip/lab-on-chip applications.