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Development of a dual specific growth rate-based fed-batch process for production of recombinant human granulocyte colony-stimulating factor in Pichia pastoris

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Abstract

A number of limitations exist for production of human granulocyte colony-stimulating factor (G-CSF) in Pichia pastoris. In this study, two different specific growth rates (0.015 h−1, 0.01 h−1) were used sequentially in the mixed substrate feeding period during methanol induction phase to enhance the G-CSF titer in the culture broth. Necessary parameters required for implementing the feeding strategy, such as specific product yield on biomass (YP/X) and maintenance coefficient (m) on glycerol, methanol, and sorbitol were estimated using continuous culture technique. Using this strategy, for the same volumetric productivity, about 20% increase in protein titer was achieved over that obtained from the run carried out at a single pre-set value of 0.015 h−1 alone. Thus, implementation of higher specific growth rate (0.015 h−1) set during initial stages of the methanol induction phase followed by a lower specific growth rate (0.01 h−1) helped in achieving increased protein titers.

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Acknowledgements

This work was supported by a grant from IIT Delhi under the `High impact research and technology leadership project’ (MI00806) awarded to SM and others.

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Author notes

  1. Ashwani Gautam

    Present address: Center for Rural Development and Technology, Indian Institute of Technology Delhi, Hauz-Khas, New Delhi, 110016, India

  2. Vikram Sahai

    Present address: Care Pro Biosciences Pvt. Ltd., I - 059, Site 5, Kasna Ind Area, Gautam Buddh Nagar, Greater Noida, UP, 201308, India

Authors and Affiliations

  1. Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz-Khas, New Delhi, 110016, India

    Saroj Mishra

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  1. Ashwani Gautam
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Correspondence to Saroj Mishra.

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Gautam, A., Sahai, V. & Mishra, S. Development of a dual specific growth rate-based fed-batch process for production of recombinant human granulocyte colony-stimulating factor in Pichia pastoris. Bioprocess Biosyst Eng 44, 103–112 (2021). https://doi.org/10.1007/s00449-020-02427-0

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