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

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. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1396–1405, 2015

[1]  Yan Zhou,et al.  Effects of peptone on hybridoma growth and monoclonal antibody formation , 2004, Cytotechnology (Dordrecht).

[2]  Harry Gruppen,et al.  Chemometric analysis of soy protein hydrolysates used in animal cell culture for IgG production - An untargeted metabolomics approach , 2014 .

[3]  L. Cantley,et al.  Understanding the Warburg Effect: The Metabolic Requirements of Cell Proliferation , 2009, Science.

[4]  Y. Schneider,et al.  Fortification of a protein-free cell culture medium with plant peptones improves cultivation and productivity of an interferon-γ-producing CHO cell line , 2003, In Vitro Cellular & Developmental Biology - Animal.

[5]  David B. Johnston,et al.  Use of Proteases to Reduce Steep Time and SO2 Requirements in a Corn Wet-Milling Process , 2001 .

[6]  A. Yoshimoto,et al.  Analysis of Structural Components and Molecular Construction of Soybean Soluble Polysaccharides by Stepwise Enzymatic Degradation , 2001, Bioscience, biotechnology, and biochemistry.

[7]  Kristine S Louis,et al.  Cell viability analysis using trypan blue: manual and automated methods. , 2011, Methods in molecular biology.

[8]  M. Choct,et al.  Soy Oligosaccharides and Soluble Non-starch Polysaccharides: A Review of Digestion, Nutritive and Anti-nutritive Effects in Pigs and Poultry , 2010 .

[9]  J. Labavitch,et al.  A SIMPLIFIED METHOD FOR ACCURATE DETERMINATION OF CELL WALL URONIDE CONTENT , 1978 .

[10]  H. Katinger,et al.  Plant Protein Hydrolysates: Preparation of Defined Peptide Fractions Promoting Growth and Production in Animal Cells Cultures , 2000, Biotechnology progress.

[11]  S. Vaughn,et al.  Characterization and antimutagenic activity of soybean saponins. , 2000, Mutation research.

[12]  H. Gruppen,et al.  Enzymatic hydrolysis as a means of expanding the cold gelation conditions of soy proteins. , 2005, Journal of Agricultural and Food Chemistry.

[13]  Gyun Min Lee,et al.  Effect of low culture temperature on specific productivity, transcription level, and heterogeneity of erythropoietin in Chinese hamster ovary cells. , 2003, Biotechnology and bioengineering.

[14]  M. Metzler,et al.  Induction of micronuclei, DNA strand breaks and HPRT mutations in cultured Chinese hamster V79 cells by the phytoestrogen coumoestrol. , 1997, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[15]  J. Vincken,et al.  Modulation of isoflavonoid composition of Rhizopus oryzae elicited soybean (Glycine max) seedlings by light and wounding. , 2013, Journal of agricultural and food chemistry.

[16]  W. Verstraete,et al.  Preparative chromatographic purification and surfactant properties of individual soyasaponins from soy hypocotyls , 2007 .

[17]  H. Katinger,et al.  Specific Effects of Synthetic Oligopeptides on Cultured Animal Cells , 2002, Biotechnology progress (Print).

[18]  Guoxiang Chen,et al.  Combined approach of NMR and chemometrics for screening peptones used in the cell culture medium for the production of a recombinant therapeutic protein. , 2007, Biotechnology and bioengineering.

[19]  Konstantin Konstantinov,et al.  The use of peptones as medium additives for the production of a recombinant therapeutic protein in high density perfusion cultures of mammalian cells , 2000, Cytotechnology.

[20]  Zhongqi Zhang,et al.  Metabolomics analysis of soy hydrolysates for the identification of productivity markers of mammalian cells for manufacturing therapeutic proteins , 2015, Biotechnology progress.

[21]  M. Corredig,et al.  Study of the role of the carbohydrate and protein moieties of soy soluble polysaccharides in their emulsifying properties. , 2004, Journal of agricultural and food chemistry.

[22]  F Gòdia,et al.  Improvement of CHO Cell Culture Medium Formulation: Simultaneous Substitution of Glucose and Glutamine , 2000, Biotechnology progress.

[23]  A. Voragen,et al.  Cell wall polysaccharides from soybean (Glycine max.) meal. Isolation and characterisation , 1998 .

[24]  H. Katinger,et al.  Enhancement of Monoclonal Antibody Production by Lysine‐Containing Peptides , 2003, Biotechnology progress.

[25]  K. Pierce,et al.  Development toward rapid and efficient screening for high performance hydrolysate lots in a recombinant monoclonal antibody manufacturing process , 2012, Biotechnology progress.

[26]  H. Englyst,et al.  Simplified method for the measurement of total non-starch polysaccharides by gas-liquid chromatography of constituent sugars as alditol acetates , 1984 .

[27]  B. Teusink,et al.  Shifts in growth strategies reflect tradeoffs in cellular economics , 2009, Molecular systems biology.

[28]  I. Koshiyama,et al.  Identification of the 7S globulin with β-conglycinin in soybean seeds , 1976 .

[29]  T. Gabryelak,et al.  Effect of the phytoestrogen, genistein‐8‐C‐glucoside, on Chinese hamster ovary cells in vitro , 2007, Cell biology international.

[30]  H. Gruppen,et al.  In vitro accessibility of untreated, toasted, and extruded soybean meals for proteases and carbohydrases , 1997 .

[31]  Jianchu Xu,et al.  The Rubber Juggernaut , 2009, Science.

[32]  F. Franek,et al.  Apoptosis and nutrition: Involvement of amino acid transport system in repression of hybridoma cell death , 2004, Cytotechnology.

[33]  Harry Gruppen,et al.  Factors causing compositional changes in soy protein hydrolysates and effects on cell culture functionality. , 2013, Journal of agricultural and food chemistry.

[34]  F. Franek Oligopeptides as External Molecular Signals Affecting Growth and Death in Animal Cell Cultures , 2008 .

[35]  D F Ollis,et al.  Enhanced Antibody Production at Slowed Growth Rates: Experimental Demonstration and a Simple Structured Model , 1990, Biotechnology progress.

[36]  M. Fussenegger,et al.  Survival Factor‐Like Activity of Small Peptides in Hybridoma and CHO Cells Cultures , 2008, Biotechnology progress.