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. 2021 Jun 11;21(7):502-513.
doi: 10.1002/elsc.202100044. eCollection 2021 Jul.

Efficacy of minute virus of mice (MVM) inactivation utilizing high temperature short time (HTST) pasteurization and suitability assessment of pasteurized, concentrated glucose feeds in Chinese hamster ovary (CHO) cell expression systems

David K Gemmell  1 , Aaron Mack  2 , Sabrina Wegmann  1 , David Han  1 , Ronald Tuccelli  1 , Matthew Johnson  1 , Corinne Miller  1
Affiliations

Efficacy of minute virus of mice (MVM) inactivation utilizing high temperature short time (HTST) pasteurization and suitability assessment of pasteurized, concentrated glucose feeds in Chinese hamster ovary (CHO) cell expression systems

David K Gemmell et al. Eng Life Sci. .

Abstract

There is a growing need to provide effective adventitious agent mitigation for high risk upstream cell culture raw materials used for the production of biologics. It is also highly important in the growing fields of cell and gene therapies. Glucose is a critical raw material necessary for effective cell growth and productivity; however, glucose is the highest risk animal-origin-free raw material for viral contamination, and often the highest risk raw material in the upstream process as more companies move to chemically defined media. This study examines the efficacy of utilizing High Temperature Short Time (HTST) pasteurization for inactivation of physiochemically resistant, worst-case parvovirus using a bench-scale HTST system. We demonstrated approximately six log inactivation of Minute Virus of Mice (MVM) in concentrated glucose feeds without impacting the subsequent performance of the glucose in a Chinese Hamster Ovary (CHO) expression system.

Keywords: high temperature short time; minute virus of mice; pasteurization; upstream cell culture risk mitigation.

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Conflict of interest statement

The authors declare no financial or commercial conflict of interest. Authors acknowledge that MilliporeSigma offer HTST‐treated glucose feeds at commercial‐scale volumes; however, this is not viewed as a conflict of interest as there are other vendors in the marketplace offering similar services. In addition, the treatment conditions recommended in this paper can be implemented by any organization on their internal HTST systems.

Figures

FIGURE 1
FIGURE 1
Minimum mean log reduction value (LRV) across all 40% (w/v) glucose runs
FIGURE 2
FIGURE 2
Minimum mean log reduction value (LRV) across All 50% (w/v) glucose runs
FIGURE 3
FIGURE 3
Comparison of viable cell density for treated and non‐treated glucose
FIGURE 4
FIGURE 4
Comparison of viability of Chinese hamster ovary (CHO) expression system with treated and non‐treated glucose
FIGURE 5
FIGURE 5
Comparison of productivity of Chinese hamster ovary (CHO) expression system with treated and non‐treated glucose
FIGURE 6
FIGURE 6
Comparison of glycan profile for treated and non‐treated glucose
FIGURE 7
FIGURE 7
Comparison of charge variant profile for treated and non‐treated glucose
See this image and copyright information in PMC

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