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A big cheese in biotherapeutics: Lactoyl leucine and isoleucine are bioavailable alternatives for canonical amino acids in cell culture media
  • +6
  • Corinna Schmidt,
  • Maria Wehsling,
  • Maxime Le Mignon,
  • Gregor Wille,
  • Yannick Rey,
  • Alisa Schnellbaecher,
  • Dmitry Zabezhinsky,
  • Markus Fischer,
  • Aline Zimmer
Corinna Schmidt
Merck KGaA
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Maria Wehsling
Merck KGaA
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Maxime Le Mignon
Merck KGaA
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Gregor Wille
Merck KGaA
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Yannick Rey
Merck KGaA
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Alisa Schnellbaecher
Merck KGaA
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Dmitry Zabezhinsky
Merck KGaA
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Markus Fischer
Merck KGaA
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Aline Zimmer
Merck KGaA
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Abstract

Increasing demands for protein-based therapeutics such as monoclonal antibodies, fusion proteins, bispecific molecules and antibody fragments require researchers to constantly find innovative solutions. To increase yields and decrease costs of next generation bioprocesses, highly concentrated cell culture media formulations are developed but often limited by the low solubility of amino acids such as tyrosine, cystine, leucine and isoleucine, in particular at physiological pH. This work sought to investigate highly soluble and bioavailable derivatives of leucine and isoleucine that are applicable for fed-batch processes. N-lactoyl-leucine and N-lactoyl-isoleucine sodium salts were tested in cell culture media and proved to be beneficial to increase the overall solubility of cell culture media formulations. These modified amino acids proved to be bioavailable for various Chinese hamster ovary (CHO) cells and were suitable for replacement of canonical amino acids in cell culture feeds. The quality of the final recombinant protein was studied in bioprocesses using the derivatives, and the mechanism of cleavage was investigated in CHO cells. Altogether, both N-lactoyl amino acids represent an advantageous alternative to canonical amino acids to develop highly concentrated cell culture media formulations to support next generation bioprocesses.

Peer review status:ACCEPTED

06 Feb 2021Submitted to Biotechnology and Bioengineering
06 Feb 2021Submission Checks Completed
06 Feb 2021Assigned to Editor
11 Feb 2021Reviewer(s) Assigned
20 Feb 2021Review(s) Completed, Editorial Evaluation Pending
20 Feb 2021Editorial Decision: Revise Minor
26 Feb 20211st Revision Received
26 Feb 2021Submission Checks Completed
26 Feb 2021Assigned to Editor
03 Mar 2021Reviewer(s) Assigned
14 Mar 2021Review(s) Completed, Editorial Evaluation Pending
14 Mar 2021Editorial Decision: Accept