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Investigation on Secondary Structure Alterations of Protein Drugs as an Indicator of Their Biological Activity Upon Thermal Exposure

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Abstract

Protein drugs are important therapeutic agents however; they may degrade during formulation processing. The objective of this study was to investigate the correlation between secondary structure alterations and the retentions of biological activity of protein upon the application of thermal stress. Catalase, horseradish peroxidase and α- chymotrypsin were employed as model proteins. Each protein was heated in a solid and solution state at a temperature of 70 °C for 1 h. Attenuated total reflectance Fourier transform infrared spectroscopy, size-exclusion chromatography and biological activity assay were performed. Results showed that heat-exposure of protein solids at 70 °C caused minimum changes in secondary structure and biological activity was almost retained. However, thermal exposure of protein aqueous solution induced significant changes in the secondary structure indicated by area overlap values and caused considerable reduction in the biological activity. The changes in secondary structures were found to be in full alignment with the loss of biological activity for both protein solids as well as aqueous solutions. Catalase lost entire biological activity upon heating in the solution state. In conclusion, the findings of the present study indicate a direct correlation between protein secondary structure alterations and the retention of biological activity which can be taken into account during the development and delivery of protein drugs formulations.

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Acknowledgements

The authors would like to thank University of Otago, Dunedin, New Zealand for providing the funding that made this study possible.

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Authors and Affiliations

  1. School of Pharmacy, International Medical University (IMU), Kuala Lumpur, Malaysia

    Farrukh Zeeshan

  2. School of Pharmacy, University of Otago, Dunedin, New Zealand

    Farrukh Zeeshan

  3. Department of Chemistry, Faculty of Science, University of Malaya (UM), Kuala Lumpur, Malaysia

    Misbah Tabbassum

  4. Department of Pharmaceutics, School of Pharmaceutical Education and Research, Jamia Hamdard, New Delhi, 110062, India

    Prashant Kesharwani

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  1. Farrukh Zeeshan
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  2. Misbah Tabbassum
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Correspondence to Prashant Kesharwani.

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Zeeshan, F., Tabbassum, M. & Kesharwani, P. Investigation on Secondary Structure Alterations of Protein Drugs as an Indicator of Their Biological Activity Upon Thermal Exposure. Protein J 38, 551–564 (2019). https://doi.org/10.1007/s10930-019-09837-4

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