Biotechnology products are often produced in mammalian cells grown in large bioreactors. The main objective of upstream manufacturing is to establish a bioreactor environment required for optimal cell growth to efficiently produce the target protein. The dynamic environment within the bioreactor is comprised of growth media along with cells, proteins, nutrients, metabolic waste products and metal ions. Metal ions have been shown to be increasingly important and can act as enzyme cofactors that directly influence the kinetics of biochemical reactions. They may also influence the biological production and quality attributes of therapeutic proteins, such as glycan formation and their distribution. Variations of the metal content in the cell culture media can affect the production of the biologic product as well as its critical quality attributes (CQAs). The current research project will discuss the application of inductively coupled plasma-mass spectrometry (ICP-MS) coupled with an advanced autosampler platform which facilitates sample and standard preparation for an automated high throughput platform that requires only a small amount of sample (&lt;1.0 mL) for analysis of numerous metals simultaneously. Developed and validated methods are applied to determine metal content in the following sample matrices: (i) CDM4CHO medium with CHO cell culture. (ii) Selective spiking of copper and zinc during CHO cell culture in bioreactors. (iii) Determination of trace metal ions in the cells during a 7-day bioreactor run in a laboratory scale parallel stirred tank bioreactor, and, (iv) Trace metal trends in a polymer bag Wave bioreactor. Trends will be discussed for each category and the importance of near-line metal analysis for monitoring bioreactor production of therapeutic proteins.

Automated and high throughput ICP-MS sample prep platform to study the effect of trace metals in upstream manufacturing of therapeutic proteins

2019 Spring National Meeting

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