Characterization of chromatographic yeast extract fractions promoting CHO cell growth

Mosser, Mathilde; Kapel, Romain; Aymes, Arnaud; Bonanno, Laurent-Michel; Olmos, Éric; Besançon, Iris; Druaux, Dominique; Chevalot, Isabelle; Marc, Ivan; Marc, Annie

doi:10.1186/1753-6561-5-s8-p99

Cited by 9 publications

(8 citation statements)

References 8 publications

(10 reference statements)

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“…And low molecular weight protein fractions of yeastolate enhanced the growth rate and productivity of insect cells . Fractionation of yeast extract using sequential steps of anion exchange, hydrophobic interaction chromatography, and size‐exclusion chromatography yielded positively charged peptides which yielded growth equivalent to the yeast extract . Di‐ and tripeptides of Primatone RL terminating in arginine are claimed to promote growth and production of recombinant proteins with peptides ranging from 280 to 546 Da .…”

Section: Discussionmentioning

confidence: 99%

“…35 Fractionation of yeast extract using sequential steps of anion exchange, hydrophobic interaction chromatography, and size-exclusion chromatography yielded positively charged peptides which yielded growth equivalent to the yeast extract. 70 Di-and tripeptides of Primatone RL terminating in arginine are claimed to promote growth and production of recombinant proteins with peptides ranging from 280 to 546 Da. 71 Although we did not test the Fraction 5 of Primatone RL which represents this size range, the Yeast 1 fraction 6 (460-570 Da) within this size range contained very low activity.…”

Section: Discussionmentioning

confidence: 99%

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The bioactivity and fractionation of peptide hydrolysates in cultures of CHO cells

Spearman

Lodewyks

Richmond

et al. 2014

Biotechnology Progress

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Peptide hydrolysate supplements in mammalian cell cultures provide enhanced growth and productivity. The objective of this study was to compare the bioactivity of ten different commercially available hydrolysates from plant, microbial, and animal sources. The peptide hydrolysates were tested as supplements to cultures of Chinese hamster ovary (CHO) cells that produce human beta interferon (β-IFN). A soy hydrolysate was shown to support high cell growth but not protein productivity compared to an animal component hydrolysate (Primatone RL). On the other hand, a yeast hydrolysate showed lower cell growth, but comparable productivity of the recombinant protein. Glycosylation analysis showed that the glycan profile of β-IFN produced in yeast hydrolysate supplemented cultures was equivalent to that from Primatone RL-supplemented cultures. Fractionation of the yeast hydrolysate and Primatone RL produced a similar protein-assayed pattern except for one extra peak at around 1 kDa in the Primatone RL profile. A fraction taken at a molecular weight range of 1.5-1.7 kDa showed the highest growth promoting activity in both samples. However, four other fractions in yeast hydrolysate and two in Primatone RL at lower molecular weights showed some growth promoting activity. In conclusion, the yeast hydrolysates provided a good alternative to the animal sourced Primatone RL for high productivity of β-IFN from CHO cells with equivalent glycosylation.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

The bioactivity and fractionation of peptide hydrolysates in cultures of CHO cells

Spearman

Lodewyks

Richmond

et al. 2014

Biotechnology Progress

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“…On one hand, peptide utilization in YE naturally depends on the composition of the peptide fraction, which precisely constitutes the main hurdle to this study due to the abundance and diversity of peptides composing these substrates. To the best of our knowledge, the YE peptide fraction has only been characterized up to now via filtration or chromatographic fractionation strategies (Mosser et al, 2011, 2015; Spearman et al, 2014, 2016), or by analyzing the composition in peptide-bound amino acids (Kevvai et al, 2014, 2016). Nonetheless, the exact nature of the peptides composing YEs still remains elusive, these compounds are therefore commonly considered as black boxes containing a complex mixture of undefined peptides.…”

Section: Introductionmentioning

confidence: 99%

Insights Into the Complexity of Yeast Extract Peptides and Their Utilization by Streptococcus thermophilus

et al. 2019

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Streptococcus thermophilus , an extensively used lactic starter, is generally produced in yeast extract-based media containing a complex mixture of peptides whose exact composition remains elusive. In this work, we aimed at investigating the peptide content of a commercial yeast extract (YE) and identifying dynamics of peptide utilization during the growth of the industrial S. thermophilus N4L strain, cultivated in 1 l bioreactors under pH-regulation. To reach that goal, we set up a complete analytical workflow based on mass spectrometry (peptidomics). About 4,600 different oligopeptides ranging from 6 to more than 30 amino acids in length were identified during the time-course of the experiment. Due to the low spectral abundance of individual peptides, we performed a clustering approach to decipher the rules of peptide utilization during fermentation. The physicochemical characteristics of consumed peptides perfectly matched the known affinities of the oligopeptide transport system of S. thermophilus . Moreover, by analyzing such a large number of peptides, we were able to establish that peptide net charge is the major factor for oligopeptide transport in S. thermophilus N4L.

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“…[7,8] Yeast extract is from a non-animal source and, as such, offers the additional benefit of increased safety from adventitious agents, which is of particular importance in the production of human therapeutics. [9] The improvement of cell growth using hydrolysates has been demonstrated in multiple cell lines (eg, CHO, [10,11] Vero, [12] and Sf-9 [13] ).…”

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confidence: 99%

Enhanced characterization of yeast hydrolysate combining acid digestion and 1D‐1H NMR targeted profiling

2021

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Yeast extract, or autolysate, is a required component for many cell-culture media, but its exact constituents and benefits are unknown. Yeast extract contains a diverse assortment of metabolites, often present in complex forms (eg, polypeptides and polynucleotides). This study employs one-dimensional proton nuclear magnetic resonance (1D-1H NMR) spectroscopy to analyze free (ie, readily available) components present in commercially available yeast autolysate. The product is monitored while further subjected to acid hydrolysis, allowing for a more robust understanding of the exact components present, particularly those contained in complex forms. The amino acids and glucose compounds behaved as expected based on other acid hydrolysis studies, and were modelled similarly. Parameter estimation was in strong agreement with pre-hydrolysis targeted 1D-1H NMR profiling. This analysis was expanded to components not as thoroughly investigated and was especially applicable to nucleic compounds. Acid hydrolysis revealed that the yeast extract was approximately 5.4% nucleic material by weight, mostly composed of adenosine, and largely provided by RNA due to the presence of uracil and lack of thymidine. Choline-containing compounds were also positively identified with an observable increase of free choline during hydrolysis. 1D-1H NMR spectroscopy allowed for the simultaneous monitoring of a significant number of yeastextract metabolites, some of which were previously unreported. Utilizing 1D-1H NMR spectroscopy in conjunction with acid hydrolysis led to a more complete view of the compounds present, and accounted for an additional 24% of the yeast-extract mass.

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Characterization of chromatographic yeast extract fractions promoting CHO cell growth

Cited by 9 publications

References 8 publications

The bioactivity and fractionation of peptide hydrolysates in cultures of CHO cells

The bioactivity and fractionation of peptide hydrolysates in cultures of CHO cells

Insights Into the Complexity of Yeast Extract Peptides and Their Utilization by Streptococcus thermophilus

Enhanced characterization of yeast hydrolysate combining acid digestion and 1D‐1H NMR targeted profiling

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