Skip to main content
SpringerLink
Log in

The Principles of Freeze-Drying and Application of Analytical Technologies

  • Protocol
  • First Online:
Cryopreservation and Freeze-Drying Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2180))

Abstract

Freeze-drying is a complex process despite the relatively small number of steps involved, since the freezing, sublimation, desorption, and reconstitution processes all play a part in determining the success or otherwise of the final product qualities, and each stage can impose different stresses on a product. This is particularly the case with many fragile biological samples, which require great care in the selection of formulation additives such as protective agents and other stabilizers. Despite this, the process is widely used, not least because once any such processing stresses can be overcome, the result is typically a significantly more stable product than was the case with the starting material. Indeed, lyophilization may be considered a gentler method than conventional air-drying methods, which tend to apply heat to the product rather than starting by removing heat as is the case here. Additionally, due to the high surface area to volume ratio, freeze-dried materials tend to be drier than their conventionally dried counterparts and also rehydrate more rapidly. This chapter provides an overview of freeze-drying (lyophilization) of biological specimens with particular reference to the importance of formulation development, characterization, and cycle development factors necessary for the commercial exploitation of freeze-dried products, and reviews the recent developments in analytical methods which have come to underpin modern freeze-drying practice.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Fanget B, Francon A (1996) A Varicella vaccine stable at 5 °C. Dev Biol Stand 87:167–171

    CAS  PubMed  Google Scholar 

  2. Adams GDJ (1995) The preservation of inocula. In: Brown MRW, Gilbert P (eds) Microbiological quality assurance: a guide towards relevance and reproducibility of inocula. CRC Press, London, pp 89–119

    Google Scholar 

  3. Hansen LJ, Daoussi R, Vervaet C, Remon JP, De Beer TR (2015) Freeze-drying of live virus vaccines: a review. Vaccine 33:5507–5519

    Article  CAS  PubMed  Google Scholar 

  4. Bindschaedler C (1999) Lyophilization process validation. In: Rey L, May JC (eds) Freeze-drying/lyophilization of pharmaceutical and biological products. Marcel Dekker, New York, pp 373–408

    Google Scholar 

  5. Liapis AI, Sadikoglu H (1997) Mathematical modeling of the primary and secondary drying stages of bulk solution freeze-drying in trays: parameter estimation and model discrimination by comparison of the theoretical results with experimental data. Dry Technol 15:791–810

    Article  Google Scholar 

  6. McAndrew TP, Hostetler D, FL DG (2019) Container and reconstitution systems for lyophilized drug products. In: Ward KR, Matejtschuk P (eds) Lyophilization of pharmaceuticals and biologicals: new technologies and approaches. Humana Press, Springer, New York, pp 193–214

    Chapter  Google Scholar 

  7. Carpenter JF, Izutsu K-I, Randolph T (2010) Freezing- and drying- induced perturbations of protein structure and mechanisms of protein protection by stabilizing additives. In: Rey L, May JC (eds) Freeze-drying/lyophilization of pharmaceutical and biological products, 3rd edn. Informa Healthcare, New York, pp 167–197

    Google Scholar 

  8. Adebayo AA, Sim-Brandenburg JW, Emmel H, Olaeye DO, Niedrig M (1998) Stability of 17D yellow fever virus vaccine using different stabilisers. Biologicals 26:309–316

    Article  CAS  PubMed  Google Scholar 

  9. Matejtschuk P, Malik K, Duru C (2019) Formulation and process development for lyophilized biological reference materials. In: Ward KR, Matejtschuk P (eds) Lyophilization of pharmaceuticals and biologicals: new technologies and approaches. Humana Press, Springer, New York, pp 33–55

    Chapter  Google Scholar 

  10. Searles JA (2010) Freezing and annealing phenomena in lyophilization. In: Rey L, May JC (eds) Freeze-drying/lyophilization of pharmaceutical and biological products, 3rd edn. Informa Healthcare, New York, pp 52–81

    Google Scholar 

  11. Ward KR, Matejtschuk P (2019) Characterization of formulations for freeze drying. In: Ward KR, Matejtschuk P (eds) Lyophilization of pharmaceuticals and biologicals: new technologies and approaches. Humana Press, Springer, New York, pp 1–32

    Chapter  Google Scholar 

  12. Luoma J, Magill G, Kumar L, Yusoff Z (2019) Controlled ice nucleation using ControLyo® pressurization-depressurization method. In: Ward KR, Matejtschuk P (eds) Lyophilization of pharmaceuticals and biologicals: new technologies and approaches. Humana Press, Springer, New York, pp 57–77

    Chapter  Google Scholar 

  13. Pisano (2019) Alternative methods of controlling nucleation in freeze drying. In: Ward KR, Matejtschuk P (eds) Lyophilization of pharmaceuticals and biologicals: new technologies and approaches. Humana Press, Springer, New York, pp 79–111

    Chapter  Google Scholar 

  14. Bhatnagar BS, Pikal MJ, Bogner RH (2007) Study of the individual contributions of ice formation and freeze-concentration on isothermal stability of lactate dehydrogenase during freezing. J Pharm Sci 97:798–814

    Article  CAS  Google Scholar 

  15. Awotwe-Otoo D, Agarabi C, Read EK, Lute S, Brorson KA, Khan MA, Shah RB (2013) Impact of controlled ice nucleation on process performance and quality attributes of a lyophilized monoclonal antibody. Int J Pharm 450:70–78

    Article  CAS  PubMed  Google Scholar 

  16. Oddone I, Arsiccio A, Duru C, Malik K, Ferguson J, Pisano R, Matejtschuk P (2019) Vacuum-induced surface freezing for the freeze-drying of the human growth hormone: how does nucleation control affect protein stability? J Pharm Sci. https://doi.org/10.1016/j.xphs.2019.04.014

  17. Geidobler R, Winter G (2013) Controlled ice nucleation in the field of freeze-drying: fundamentals and technology review. Eur J Pharm Biopharm 85:214–222

    Article  CAS  PubMed  Google Scholar 

  18. Patel SM, Nail SL, Pikal MJ, Geidobler R, Winter G, Hawe A, Davagnino J, Gupta SR (2017) Lyophilized product cake appearance: what is acceptable? J Pharm Sci 106:1706–1721

    Article  CAS  PubMed  Google Scholar 

  19. Patapoff TW, Overcashier D, Hsu C, Nguyen TH, Borchardt RT (1996) Effects of reducing sugars on the chemical stability of human relaxin in the lyophilized state. J Pharm Sci 85:873–877

    Article  PubMed  Google Scholar 

  20. Pikal MJ (1999) Mechanisms of protein stabilization during freeze-drying and storage: the relative importance of thermo stabilization and glassy state relaxation dynamics. In: Rey L, May JC (eds) Freeze-drying/lyophilization of pharmaceutical and biological products. Marcel Dekker, New York, pp 161–198

    Google Scholar 

  21. Morgan CA, Herman N, White PA, Vesey G (2006) Preservation of microrganisms by drying; a review. J Microbiol Methods 66:183–193

    Article  CAS  PubMed  Google Scholar 

  22. Rey LR (1999) Glimpses into the realm of freeze-drying: classic issues and new ventures. In: Rey L, May JC (eds) Freeze-drying/lyophilization of pharmaceutical and biological products. Marcel Dekker, New York, pp 1–30

    Google Scholar 

  23. Padilla AM, Pikal MJ (2010) Phase separation of freeze-dried amorphous solids: the occurrence and detection of multiple amorphous phases in pharmaceutical systems. In: Rey L, May JC (eds) Freeze drying/lyophilization of pharmaceuticals and biological products, 3rd edn. Informa Healthcare, New York, pp 82–111

    Google Scholar 

  24. Mockus LN, Paul TW, Pease NA, Harper NJ, Basu PK, Oslos EA, Sacha GA, Kuu WY, Hardwick LM, Karty JJ, Pikal MJ, Hee E, Khan MA, Nail SL (2011) Quality by design in formulation and process development for a freeze-dried, small molecule parenteral product: a case study. Pharm Dev Technol 16:549–576

    Article  CAS  PubMed  Google Scholar 

  25. Patel SM, Pikal MJ (2011) Emerging freeze-drying process development and scale-up issues. AAPS Pharm Sci Tech 12:372–378

    Article  Google Scholar 

  26. Ward K, Cowen A, Peacock T (2012) Freeze drying method. Patent WO/2012/098358

    Google Scholar 

  27. MacKenzie AP (1964) Apparatus for microscopic observations during freeze drying. Biodynamica 9:213–222

    CAS  PubMed  Google Scholar 

  28. Ward K, Matejtschuk P (2010) The use of microscopy, thermal analysis and impedance measurements to establish critical formulation parameters for freeze-drying cycle development. In: Rey L, May JC (eds) Freeze drying/lyophilization of pharmaceuticals and biological products, 3rd edn. Informa Healthcare, New York, pp 112–135

    Google Scholar 

  29. Franks F, Auffret A (2007) Freeze-drying of pharmaceuticals and biopharmaceuticals: principles and practice. RSC Press, Cambridge UK

    Google Scholar 

  30. Kett V, McMahon D, Ward K (2004) Freeze-drying of protein pharmaceuticals - the application of thermal analysis. Cryo Letters 25:389–404

    CAS  PubMed  Google Scholar 

  31. Mehta M, Bhardwaj SP, Suryanarayanan R (2013) Controlling the physical form of mannitol in freeze dried systems. Eur J Pharm Biopharm 85:207–213

    Article  CAS  PubMed  Google Scholar 

  32. Duddu SP, Dal Monte PR (1997) Effect of glass transition temperature on the stability of lyophilized formulations containing a chimeric therapeutic monoclonal antibody. Pharm Res 14:591–595

    Article  CAS  PubMed  Google Scholar 

  33. Wang W (2000) Lyophilization and development of solid protein pharmaceuticals. Int J Pharm 203:1–60

    Article  CAS  PubMed  Google Scholar 

  34. Grant Y, Matejtschuk P, Dalby PA (2009) Rapid optimization of protein freeze drying formulations using ultra scale down and factorial design of experiment in microplates. Biotechnol Bioeng 104:957–964

    Article  CAS  PubMed  Google Scholar 

  35. Bourles E, de Lannoy F, Scutella B, Fonseca F, Trelea IC, Passot S (2019) Scale-up of freeze drying cycles, the use of process analytical technology (PAT), and statistical analysis. In: Ward KR, Matejtschuk P (eds) Lyophilization of pharmaceuticals and biologicals: new technologies and approaches. Humana Press, Springer, New York, pp 215–240

    Chapter  Google Scholar 

  36. Cherry C (2019) Containment options for the freeze-drying of biological entities and potent materials. In: Ward KR, Matejtschuk P (eds) Lyophilization of pharmaceuticals and biologicals: new technologies and approaches. Humana Press, Springer, New York, pp 143–155

    Chapter  Google Scholar 

  37. Schneid S, Gieseler H (2008) Evaluation of a new wireless temperature remote interrogation system (TEMPRIS) to measure product temperature during freeze drying. AAPS Pharm Sci Tech 9:729–739

    Article  CAS  Google Scholar 

  38. Tang C, Nail SL, Pikal MJ (2005) Freeze-drying process design by manometric temperature measurement: design of a smart freeze-dryer. Pharm Res 22:685–700

    Article  CAS  PubMed  Google Scholar 

  39. Kessler WJ, Gong E (2019) Tunable diode laser absorption spectroscopy in lyophilization. In: Ward KR, Matejtschuk P (eds) Lyophilization of pharmaceuticals and biologicals: new technologies and approaches. Humana Press, Springer, New York, pp 113–141

    Chapter  Google Scholar 

  40. Sylvester B, Porfire A, Van Bockstal P-J, Porav S, Achim M, De Beer T, Tomuţă J (2018) Formulation of long-circulating liposomes and in-line monitoring of the freeze-drying process using a NIR spectroscopy tool. J Pharm Sci 107:139–148

    Article  CAS  PubMed  Google Scholar 

  41. Fissore D, Pisano R, Barresi AA (2018) Process analytical technology for monitoring pharmaceuticals freeze-drying—a comprehensive review. Dry Technol 36:1839–1865

    Article  Google Scholar 

  42. Cook IA, Ward KR (2011) Headspace moisture mapping and the information that can be gained about freeze-dried materials and processes. PDA J Pharm Sci Technol 65:457–467

    Article  CAS  PubMed  Google Scholar 

  43. Hedberg SHM, Devi S, Duralliu A, Williams DR (2019) Mechanical behavior and structure of freeze-dried cakes. In: Ward KR, Matejtschuk P (eds) Lyophilization of pharmaceuticals and biologicals: new technologies and approaches. Humana Press, Springer, New York, pp 327–351

    Chapter  Google Scholar 

  44. Haeuser C, Goldbach P, Huwyler J, Friess W, Allmendinger A (2018) Imaging techniques to characterize appearance of freeze-dried products. J Pharm Sci 107:2810–2822

    Article  CAS  PubMed  Google Scholar 

  45. Atotwe-Otoo D, Khan M (2019) Regulatory aspects of freeze-drying. In: Ward KR, Matejtschuk P (eds) Lyophilization of pharmaceuticals and biologicals: new technologies and approaches. Humana Press, Springer, New York, pp 173–192

    Chapter  Google Scholar 

Download references

Acknowledgments

The authors would like to acknowledge colleagues at Biopharma Process Systems and NIBSC for their input and discussion to this chapter.

Author information

Authors and Affiliations

  1. Biopharma Process Systems Ltd, Winchester, UK

    Kevin R. Ward

  2. National Institute for Biological Standards and Control (NIBSC), Potters Bar, UK

    Paul Matejtschuk

Authors
  1. Kevin R. Ward
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Paul Matejtschuk
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kevin R. Ward .

Editor information

Editors and Affiliations

  1. Unit for Reproductive Medicine—Clinic for Horses, Biostabilization Laboratory—Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, University of Veterinary Medicine Hannover, Hannover, Germany, University of Veterinary Medicine Hannover, Hannover, Germany

    Willem F. Wolkers

  2. Unit for Reproductive Medicine—Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany

    Harriëtte Oldenhof

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Ward, K.R., Matejtschuk, P. (2021). The Principles of Freeze-Drying and Application of Analytical Technologies. In: Wolkers, W.F., Oldenhof, H. (eds) Cryopreservation and Freeze-Drying Protocols. Methods in Molecular Biology, vol 2180. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0783-1_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0783-1_3

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0782-4

  • Online ISBN: 978-1-0716-0783-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation