Development and Application of a Rational Design for Evaluation and Optimization of Animal Derived Component Free Media Formulation

Date

2013-05-01

Authors

Murayyan, Abdulmonem

Journal Title

Journal ISSN

Volume Title

Publisher

University of Guelph

Abstract

Cell culture media used in the manufacture of biopharmaceuticals conventionally contain many animal derived components. These components can harbor adventitious agents which can be transmitted through biotherapeutics, employed in the medical treatment of immunocompromised patients. An ADCF (animal derived component free) medium formulation obviates this concern. A rational method for the rapid and efficient screening and optimization of ADCF media while preserving, if not enhancing, cellular growth and protein productivity is needed. CHO (Chinese Hamster Ovary) cells, widely used as a production platform in industry, expressing a recombinant protein, were employed as a model system. Design of Experiment (DOE) and statistical analysis were employed to assess the impact of media formulation on cellular physiology. Metabolic flux, cellular growth, and protein productivity were evaluated as the measures of ADCF media formulation success. Measurements of extracellular metabolites were determined by HPLC and enzymatic methods. Recombinant protein production was measured by HPLC. This research demonstrates the successful screening and optimization of four plant hydrolysate mixtures (2 soy and 2 wheat) as a replacement for animal derived components.

Description

Keywords

Mammalian Cell Culture, Design of Experiment, Animal Derived Component Free Media

Citation