Digging deeper into HEK293 metabolism: Growth inhibitory metabolic byproduct production in fed-batch cultures and their control

Digging deeper into HEK293 metabolism: Growth inhibitory metabolic byproduct production in fed-batch cultures and their control | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL Biotechnology Journal This is a preprint and has not been peer reviewed. Data may be preliminary. 19 February 2025 V1 Latest version Share on Digging deeper into HEK293 metabolism: Growth inhibitory metabolic byproduct production in fed-batch cultures and their control Authors : Cameron Harrington and Bhanu Mulukutla 0000-0002-7515-9599 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.173995947.71764891/v1 497 views 151 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract not-yet-known not-yet-known not-yet-known unknown Human Embryonic Kidney 293 (HEK293) cells are currently one of the preferred host cell lines for production of biologics, specifically, AAV−based viral vectors. These fast-growing cells consume significant amounts of nutrients and often convert them into byproducts such as lactate and ammonia. In fed-batch cultures, accumulation of lactate and ammonia to high levels can inhibit cell proliferation. In this study, we demonstrate that lactate and ammonia accumulation alone doesn’t fully explain the growth inhibition observed in HEK293 fed-batch cultures. Growth inhibition was noted even when the levels of these byproducts were well controlled. Instead, we show that several previously unknown compounds accumulate in HEK293 cell fed-batch cultures, some of which can inhibit HEK293 cell growth either individually or synergistically. Many of these newly identified compounds are intermediates or byproducts of amino acid catabolism. When residual levels of the source amino acids for these novel byproducts were controlled in the low concentration range (~1mM) in HEK293 fed-batch cultures, lactate accumulated to higher levels, causing growth inhibition. This prompted the need for employing HIPDOG control strategy for lactate management. In HIPDOG cultures, controlling the source amino acids at low concentrations resulted in lower accumulations of the corresponding growth-inhibitory byproducts when compared to the control HIPDOG conditions with typical levels of amino acids. This led to higher viable cell densities (VCD) and viabilities in low amino acid conditions. Strategies that reduce byproduct accumulation, whether classical or novel byproducts, in HEK293 fed-batch processes can result in enhanced VCDs potentially leading to higher volumetric productivities Supplementary Material File (manuscript_harrington_mulukutla.docx) Download 192.70 KB Information & Authors Information Version history V1 Version 1 19 February 2025 Copyright This work is licensed under a Non Exclusive No Reuse License. Collection Biotechnology Journal Keywords bioprocess engineering hek293 cells low amino acid process mammalian cells metabolic byproducts metabolism metabolomics Authors Affiliations Cameron Harrington Pfizer Andover View all articles by this author Bhanu Mulukutla 0000-0002-7515-9599 [email protected] Pfizer Andover View all articles by this author Metrics & Citations Metrics Article Usage 497 views 151 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Cameron Harrington, Bhanu Mulukutla. Digging deeper into HEK293 metabolism: Growth inhibitory metabolic byproduct production in fed-batch cultures and their control. Authorea . 19 February 2025. DOI: https://doi.org/10.22541/au.173995947.71764891/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. 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