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The role of protein hydrolysates in prolonging viability and enhancing antibody production of CHO cells

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Abstract

Four independent mAb-producing CHO cell lines were grown in media supplemented with one of seven protein hydrolysates of animal and plant origin. This generated a 7x4 matrix of replicate cultures which was analysed for viable cell density and mAb productivity. In all cultures, a consistent growth rate was shown in batch culture up to 4 to 5 days. Differences between cultures appeared in the decline phase which was followed up to 7 days beyond the start of the cultures. There was a marginal but significant overall increase (x1.1) in the integral viable cell density (IVCD) in the presence of hydrolysate but a more substantial increase in the cell-specific mAb (qMab) productivity (x1.5). There were individual differences between hydrolysates in terms of enhancement of mAb productivity, the highest being a 166% increase of mAb titre (to 117 mg/L) in batch cultures of CHO-EG2 supplemented with UPcotton hydrolysate. The effect of one of the most active hydrolysates (HP7504) on antibody glycosylation was investigated. This showed no change in the predominant seven glycans produced but a significant increase in the galactosylation and sialylation of some but not all the antibodies. Overall, the animal hydrolysate, Primatone and two cotton-derived hydrolysates provided the most substantial benefit for enhanced productivity. The cotton-based hydrolysates can be viewed as valuable supplements for animal-derived component-free (ADCF) media and as a source for the investigation of chemically defined bioactive components.

Key points

• Protein hydrolysates enhanced both IVCD & qMab; the effect on qMab being consistently greater.

• Cotton-based hydrolysates showed high bioactivity and potential for use in serum-free media.

• Enhanced galactosylation and sialylation was shown for some of the Mabs tested.

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Data availability

Glycan structures were validated by reference to the Glycostore database (www.glycostore.org).

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Acknowledgements

The work was supported by an Enterprise Ireland, Innovation Partnership Project grant IP-2018-0717 in conjunction with the Kerry Group. We thank Kyle Liu and Hans Huttinga of Kerry for insightful comments on the manuscript.

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Authors and Affiliations

  1. Cell Technology Group, National Institute for Bioprocessing, Research and Training (NIBRT), Fosters Avenue, Mount Merrion, Blackrock, Dublin, A94 X099, Ireland

    Ismael Obaidi, Letícia Martins Mota, Andrew Quigley & Michael Butler

  2. College of Pharmacy, University of Babylon, Babylon, Iraq

    Ismael Obaidi

  3. School of Chemical and Bioprocess Engineering, University College Dublin, Dublin, D04 V1W8, Ireland

    Letícia Martins Mota & Michael Butler

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  1. Ismael Obaidi
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IO designed and conducted the experiments as well as writing the manuscript. AQ designed and conducted the experiments. LMM performed glycan analysis of the Mabs. MB conceived the research and edited and approved the manuscript.

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Correspondence to Michael Butler.

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Obaidi, I., Mota, L.M., Quigley, A. et al. The role of protein hydrolysates in prolonging viability and enhancing antibody production of CHO cells. Appl Microbiol Biotechnol 105, 3115–3129 (2021). https://doi.org/10.1007/s00253-021-11244-8

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