Chemometric analysis of soy protein hydrolysates used in animal cell culture for IgG production – An untargeted metabolomics approach

Gupta, Abhishek; Hageman, Jos A.; Wierenga, Peter A.; Boots, Jan-Willem P.; Gruppen, Harry

doi:10.1016/j.procbio.2013.10.013

Cited by 24 publications

(21 citation statements)

References 34 publications

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“…The identified productivity markers, however, are not universally applicable to all CHO cell lines, as indicated by the difference in the two cell lines used in this study. A recent study also reported an entirely different set of key components in soy hydrolysates toward a CRL‐11397 CHO cell line producing a mAb …”

Section: Discussionmentioning

confidence: 97%

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Metabolomics analysis of soy hydrolysates for the identification of productivity markers of mammalian cells for manufacturing therapeutic proteins

Richardson

Shah

Bondarenko

et al. 2015

Biotechnology Progress

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Soy hydrolysates are widely used as a nutrient supplement in mammalian cell culture for the production of recombinant proteins. The batch-to-batch variability of a soy hydrolysate often leads to productivity differences. This report describes our metabolomics platform, which includes a battery of LC-MS/MS modes of operation, and advanced data analysis software for automated data processing. The platform was successfully used for screening productivity markers in soy hydrolysates during the production of two therapeutic antibodies in two Chinese hamster ovary cell lines. A total of 123 soy hydrolysate batches were analyzed, from which 62 batches were used in the production runs of cell line #1 and 12 batches were used in the production runs of cell line #2. For cell line #1, out of 19 amino acids, 106 other metabolites and 4,131 peptides identified in the soy hydrolysate batches being used, several nucleosides and short hydrophobic peptides showed negative correlation with antibody titer, while ornithine, citrulline and several amino acids and organic acids correlated positively with titer. For cell line #2, only ornithine and citrulline showed strong positive correlation. When ornithine was spiked into the culture media, both cell lines demonstrated accelerated cell growth, indicating ornithine as a root cause of the performance difference. It is proposed that better soy hydrolysate performance resulted from better bacterial fermentation during the hydrolysate production. A few selected markers were used to predict the performance of other soy hydrolysate batches for cell line #1. The predicted titers agreed with the experimental values with good accuracy.

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Section: Discussionmentioning

confidence: 97%

“…In a nontargeted approach, correlation analysis is performed on all identified compounds, before any hypotheses are made. A similar methodology has also been used in a recent report to find key components in soy hydrolysates that is important for the production of a mAb using a CRL‐11397 Chinese hamster ovary (CHO) cell line …”

Section: Introductionmentioning

confidence: 99%

Metabolomics analysis of soy hydrolysates for the identification of productivity markers of mammalian cells for manufacturing therapeutic proteins

Richardson

Shah

Bondarenko

et al. 2015

Biotechnology Progress

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“…Considering the obvious advantages of hydrolysates, some efforts had been paid on the identification and purification of the effective molecules from the raw hydrolysates. However, most works just provided statistical relevance between some compounds and product concentrations (Gupta et al 2014;Richardson et al 2015) or separated a fraction with less components (Michiels et al 2011;Mosser et al 2012). In the present study, it was found that YE improved Fc-fusion protein production in multiple aspects.…”

Section: Discussionmentioning

confidence: 99%

Understanding the intracellular effects of yeast extract on the enhancement of Fc-fusion protein production in Chinese hamster ovary cell culture

Sun

Liu

et al. 2015

Appl Microbiol Biotechnol

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Yeast extract (YE), as a non-animal source additive for mammalian cell culture medium, has been widely used for manufacturing of therapeutic proteins. In the present study, one particular YE was found to have significantly improved the specific productivity (q p) of Fc-fusion protein in recombinant Chinese hamster ovary (rCHO) cell culture. In order to elucidate the intracellular effects of YE on protein productivity, steps of the target protein synthesis process were investigated to unveil their variations caused by YE addition. Stepwise analysis on Fc-fusion protein synthesis process showed that YE enhanced Fc-fusion protein gene transcription with cell cycle arrest at G1 phase; mammalian target of rapamycin (mTOR) signaling pathway was activated to enhance the translation of Fc-fusion protein, and the block in post-translational steps of Fc-fusion protein was alleviated by YE addition as well. Our results revealed the responses of multiple protein production steps to the addition of YE and provided a practical guidance for the separation and application of active compounds from hydrolysates.

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“…2 Phenyllactate, lactate, trigonelline, ferulic acid, chiro-inositol, and adenine were shown to explain part of the variability in IVCD (20-29%) and IgG production (22-30%). 2 It is important to note that the effect of hydrolysate supplementation depends on the cells as well as on the CD medium used in the experiment. As a result, the exact contribution of the different compounds or compound classes in the CD medium or the supplemented hydrolysates is difficult to determine.…”

Section: Introductionmentioning

confidence: 95%

“…The total IgG production (mg mL 21 ) was measured using an enzyme-linked immunosorbent assay. 2 The specific IgG production (fgÁcell 21 day…”

Section: Cell Culturementioning

confidence: 99%

Influence of protein and carbohydrate contents of soy protein hydrolysates on cell density and IgG production in animal cell cultures

Gupta

Wierenga

Gruppen

et al. 2015

Biotechnol Progress

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The variety of compounds present in chemically defined media as well as media supplements makes it difficult to use a mechanistic approach to study the effect of supplement composition on culture functionality. Typical supplements, such as soy protein hydrolysates contain peptides, amino acids, carbohydrates, isoflavones, and saponins. To study the relative contribution of these compound classes, a set of hydrolysates were produced, containing 58-83% proteinaceous material and 5-21% carbohydrates. While the content of the different compounds classes varied, the composition (e.g., peptide profiles, carbohydrate composition) did not vary in hydrolysates. The hydrolysates were supplemented to a chemically defined medium in cell culture, based on equal weight and on equal protein levels. The latter showed that an increase in the carbohydrate concentration significantly (P value < 0.004) increased integral viable cell density (IVCD) (R = 0.7) and decreased total IgG (R = -0.7) and specific IgG production (R = -0.9). The extrapolation of effects of protein concentration showed that an increase in protein concentration increased total and specific IgG production and suppressed IVCD. In addition to proteins and carbohydrates, the functionality of soy protein hydrolysates may be modulated by the presence of other minor compounds. In the current study, the large differences in the balance between total proteins and total carbohydrates in the supplemented media seem to be a main factor influencing the balance between the viable cell density, total IgG, and specific IgG production.

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Chemometric analysis of soy protein hydrolysates used in animal cell culture for IgG production – An untargeted metabolomics approach

Cited by 24 publications

References 34 publications

Metabolomics analysis of soy hydrolysates for the identification of productivity markers of mammalian cells for manufacturing therapeutic proteins

Metabolomics analysis of soy hydrolysates for the identification of productivity markers of mammalian cells for manufacturing therapeutic proteins

Understanding the intracellular effects of yeast extract on the enhancement of Fc-fusion protein production in Chinese hamster ovary cell culture

Influence of protein and carbohydrate contents of soy protein hydrolysates on cell density and IgG production in animal cell cultures

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