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High-Throughput Process Development: I. Process Chromatography

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Protein Downstream Processing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1129))

Abstract

Chromatographic separation serves as “a workhorse” for downstream process development and plays a key role in removal of product-related, host cell-related, and process-related impurities. Complex and poorly characterized raw materials and feed material, low feed concentration, product instability, and poor mechanistic understanding of the processes are some of the critical challenges that are faced during development of a chromatographic step. Traditional process development is performed as trial-and-error-based evaluation and often leads to a suboptimal process. High-throughput process development (HTPD) platform involves an integration of miniaturization, automation, and parallelization and provides a systematic approach for time- and resource-efficient chromatography process development. Creation of such platforms requires integration of mechanistic knowledge of the process with various statistical tools for data analysis. The relevance of such a platform is high in view of the constraints with respect to time and resources that the biopharma industry faces today. This protocol describes the steps involved in performing HTPD of process chromatography step. It described operation of a commercially available device (PreDictor™ plates from GE Healthcare). This device is available in 96-well format with 2 or 6 μL well size. We also discuss the challenges that one faces when performing such experiments as well as possible solutions to alleviate them. Besides describing the operation of the device, the protocol also presents an approach for statistical analysis of the data that is gathered from such a platform. A case study involving use of the protocol for examining ion-exchange chromatography of granulocyte colony-stimulating factor (GCSF), a therapeutic product, is briefly discussed. This is intended to demonstrate the usefulness of this protocol in generating data that is representative of the data obtained at the traditional lab scale. The agreement in the data is indeed very significant (regression coefficient 0.93). We think that this protocol will be of significant value to those involved in performing high-throughput process development of process chromatography.

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Acknowledgements

Financial assistance from the Department of Biotechnology, Government of India, New Delhi, is gratefully acknowledged. We are also thankful to GE Healthcare (Uppsala, Sweden) for providing us with financial support and some of the consumable items used in this investigation. We also thank Dionex Corporation, USA, for donating some of the equipment for this project.

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Authors and Affiliations

  1. Department of Chemical Engineering, Indian Institute of Technology, Hauz Khas, New Delhi, 110016, India

    Anurag S. Rathore

  2. Department of Chemical Engineering, Indian Institute of Technology, New Delhi, India

    Rahul Bhambure

Authors
  1. Anurag S. Rathore
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  2. Rahul Bhambure
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Corresponding author

Correspondence to Anurag S. Rathore .

Editor information

Editors and Affiliations

  1. Laboratory of Enzyme Technology, Agricultural University of Athens Department of Agr. Biotechnology, Athens, Greece

    Nikolaos E. Labrou

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Rathore, A.S., Bhambure, R. (2014). High-Throughput Process Development: I. Process Chromatography. In: Labrou, N. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 1129. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-977-2_3

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  • DOI: https://doi.org/10.1007/978-1-62703-977-2_3

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-976-5

  • Online ISBN: 978-1-62703-977-2

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