Date of Award

Spring 5-7-2021

Level of Access Assigned by Author

Open-Access Thesis

Degree Name

Master of Science in Biomedical Engineering

Department

Chemical Engineering

Advisor

G. Peter Van Walsum

Second Committee Member

Karissa Tilbury

Third Committee Member

José A. Fernández Robledo

Abstract

Perkinsus marinus is a marine protozoan responsible for “Dermo” disease in the eastern oyster species, Crassostrea virginica. P. marinus has been detected along the Atlantic Coast of the United States and Mexico. In laboratory studies, many parasites need to be maintained in vivo, which complicates the study of the organism. However, In vitro cell culture for P. marinus at a scale of 1 mL was established in 1993 by several groups with the 1995 optimization (Gauthier & Vasta, 1993, 1995) considered the gold standard for small scale growth of this species. In addition to its importance as a parasite of an important food source, P. marinus is notable among easily cultured microbes in its ability to produce, fold and excrete very large, complex proteins. Such proteins could potentially be used for therapeutic applications, such as development of vaccines.

In order to be useful as an organism to produce vaccines, we need to learn how to grow the organism at larger scale. Transitioning the growth procedures from small culture vials to tenfold larger bioreactors is a first step towards achieving this scale up. Growing P. marinus in a bioreactor introduces new production variables and challenges, such as: aeration, pH control, temperature control, and cost of media. In this study we developed methods and procedures to grow P. marinus in bioreactors and have also developed a lower cost growth medium that reduced the cost of growth medium ingredients by about 60%. Higher order cell culture comes with expensive media components, most notably fetal bovine serum (FBS), which provides essential growth factors and cytokines for growing cells. A promising replacement for FBS was found to be chicken serum (CS). It has the potential to provide growth factors and cytokines at a much lower cost than FBS.

SuperPro Designer v10 (Intelligen, Inc., Scotch Plains, New Jersey) is a process simulator software designed to analyze the techno-economics of commercial-scale bioprocesses. SuperPro Designer v10 was used to translate the experimental fermentation data from this study into a process model that provides insight into the financial feasibility of growing P. marinus at an industry scale.

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