AAPS PharmSciTech

, Volume 19, Issue 1, pp 448–459 | Cite as

Application of Optical Coherence Tomography Freeze-Drying Microscopy for Designing Lyophilization Process and Its Impact on Process Efficiency and Product Quality

  • Maxwell Korang-YeboahEmail author
  • Charudharshini Srinivasan
  • Akhtar Siddiqui
  • David Awotwe-Otoo
  • Celia N. Cruz
  • Ashraf MuhammadEmail author
Research Article


Optical coherence tomography freeze-drying microscopy (OCT-FDM) is a novel technique that allows the three-dimensional imaging of a drug product during the entire lyophilization process. OCT-FDM consists of a single-vial freeze dryer (SVFD) affixed with an optical coherence tomography (OCT) imaging system. Unlike the conventional techniques, such as modulated differential scanning calorimetry (mDSC) and light transmission freeze-drying microscopy, used for predicting the product collapse temperature (Tc), the OCT-FDM approach seeks to mimic the actual product and process conditions during the lyophilization process. However, there is limited understanding on the application of this emerging technique to the design of the lyophilization process. In this study, we investigated the suitability of OCT-FDM technique in designing a lyophilization process. Moreover, we compared the product quality attributes of the resulting lyophilized product manufactured using Tc, a critical process control parameter, as determined by OCT-FDM versus as estimated by mDSC. OCT-FDM analysis revealed the absence of collapse even for the low protein concentration (5 mg/ml) and low solid content formulation (1%w/v) studied. This was confirmed by lab scale lyophilization. In addition, lyophilization cycles designed using Tc values obtained from OCT-FDM were more efficient with higher sublimation rate and mass flux than the conventional cycles, since drying was conducted at higher shelf temperature. Finally, the quality attributes of the products lyophilized using Tc determined by OCT-FDM and mDSC were similar, and product shrinkage and cracks were observed in all the batches of freeze-dried products irrespective of the technique employed in predicting Tc.


lyophilization optical coherence freeze-drying microscopy collapse temperature glass transition temperature protein formulation 



The authors would like to thank Dr. Haiou Qu and Dr. Yifan Wang, ORISE Fellows at the Food and Drug Administration, for their contribution.

Compliance with Ethical Standards


The opinions expressed in this work are only of the author and should not be construed to represent FDA’s views or policies.

Supplementary material


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  1. 1.
    LaTorre-Snyder M. Lyophilization: The basics. Pharmaceutical Processing. 2017.
  2. 2.
    Werk T, Ludwig IS, Luemkemann J, Mahler H-C, Huwyler J, Hafner M. Technology, applications, and process challenges of dual chamber systems. J Pharm Sci. 2016;105(1):4–9.CrossRefPubMedGoogle Scholar
  3. 3.
    Rambhatla S, Obert JP, Luthra S, Bhugra C, Pikal MJ. Cake shrinkage during freeze drying: a combined experimental and theoretical study. Pharm Dev Technol. 2005;10(1):33–40.CrossRefPubMedGoogle Scholar
  4. 4.
    Bellows RJ, King CJ. Freeze-drying of aqueous solutions: maximum allowable operating temperature. Cryobiology. 1972;9(6):559–61.CrossRefPubMedGoogle Scholar
  5. 5.
    Her LM, Nail SL. Measurement of glass transition temperatures of freeze-concentrated solutes by differential scanning calorimetry. Pharm Res. 1994;11(1):54–9.CrossRefPubMedGoogle Scholar
  6. 6.
    Adams GD, Irons LI. Some implications of structural collapse during freeze-drying using Erwinia caratovora L-asparaginase as a model. J Chem Technol Biotechnol. 1993;58(1):71–6.CrossRefPubMedGoogle Scholar
  7. 7.
    Passot S, Fonseca F, Barbouche N, Marin M, Alarcon-Lorca M, Rolland D, et al. Effect of product temperature during primary drying on the long-term stability of lyophilized proteins. Pharm Dev Technol. 2007;12(6):543–53.CrossRefPubMedGoogle Scholar
  8. 8.
    Chang BS, Beauvais RM, Dong A, Carpenter JF. Physical factors affecting the storage stability of freeze-dried interleukin-1 receptor antagonist: glass transition and protein conformation. Arch Biochem Biophys. 1996;331(2):249–58.CrossRefPubMedGoogle Scholar
  9. 9.
    Tang X, Pikal MJ. Design of freeze-drying processes for pharmaceuticals: practical advice. Pharm Res. 2004;21(2):191–200.CrossRefPubMedGoogle Scholar
  10. 10.
    Carpenter JF, Pikal MJ, Chang BS, Randolph TW. Rational design of stable lyophilized protein formulations: some practical advice. Pharm Res. 1997;14(8):969–75.CrossRefPubMedGoogle Scholar
  11. 11.
    MacKenzie AP. Collapse during freeze drying—qualitative and quantitative aspects. In: Goldblith SA, Rey L, Rothmayr WW, editors. Freeze Drying and Advanced Food Technology. London: Academic Press. 1975. P. 277–307.Google Scholar
  12. 12.
    Pikal MJ. Use of laboratory data in freeze drying process design: heat and mass transfer coefficients and the computer simulation of freeze drying. J Parenter Sci Technol Publ Parenter Drug Assoc. 1985;39(3):115–39.Google Scholar
  13. 13.
    Meister E, Gieseler H. Freeze-dry microscopy of protein/sugar mixtures: drying behavior, interpretation of collapse temperatures and a comparison to corresponding glass transition data. J Pharm Sci. 2009;98(9):3072–87.CrossRefPubMedGoogle Scholar
  14. 14.
    Pikal MJ, Shah S. The collapse temperature in freeze-drying—dependence on measurement methodology and rate of water removal from the glassy phase. Int J Pharm. 1990;62(2–3):165–86.CrossRefGoogle Scholar
  15. 15.
    Colandene JD, Maldonado LM, Creagh AT, Vrettos JS, Goad KG, Spitznagel TM. Lyophilization cycle development for a high-concentration monoclonal antibody formulation lacking a crystalline bulking agent. J Pharm Sci. 2007;96(6):1598–608.CrossRefPubMedGoogle Scholar
  16. 16.
    Johnson RLL. Freeze-drying protein formulations above their collapse temperatures: possible issues and concerns. Am Pharm Rev. 2011;14(3):50–4.Google Scholar
  17. 17.
    Johnson RE, Oldroyd ME, Ahmed SS, Gieseler H, Lewis LM. Use of manometric temperature measurements (MTM) to characterize the freeze-drying behavior of amorphous protein formulations. J Pharm Sci. 2010;99(6):2863–73.CrossRefPubMedGoogle Scholar
  18. 18.
    Greco K, Mujat M, Galbally-Kinney KL, Hammer DX, Ferguson RD, Iftimia N, et al. Accurate prediction of collapse temperature using optical coherence tomography-based freeze-drying microscopy. J Pharm Sci. 2013;102(6):1773–85.Google Scholar
  19. 19.
    Mujat M, Greco K, Galbally-Kinney KL, Hammer DX, Ferguson RD, Iftimia N, et al. Optical coherence tomography-based freeze-drying microscopy. Biomed Opt Express. 2012;3(1):55–63.Google Scholar
  20. 20.
    Srinivasan C, Siddiqui A, Korang-Yeboah M, Khan MA. Stability characterization and appearance of particulates in a lyophilized formulation of a model peptide hormone-human secretin. Int J Pharm. 2015;481(1–2):104–13.CrossRefPubMedGoogle Scholar
  21. 21.
    Ullrich S, Seyferth S, Lee G. Measurement of shrinkage and cracking in lyophilized amorphous cakes. Part I: final-product assessment. J Pharm Sci. 2015;104(1):155–64.CrossRefPubMedGoogle Scholar
  22. 22.
    Militello V, Casarino C, Emanuele A, Giostra A, Pullara F, Leone M. Aggregation kinetics of bovine serum albumin studied by FTIR spectroscopy and light scattering. Biophys Chem. 2004;107(2):175–87.CrossRefPubMedGoogle Scholar
  23. 23.
    Awotwe-Otoo D, Agarabi C, Keire D, Lee S, Raw A, Yu L, et al. Physicochemical characterization of complex drug substances: evaluation of structural similarities and differences of protamine sulfate from various sources. AAPS J. 2012;14(3):619–26.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Kelly SM, Price NC. The application of circular dichroism to studies of protein folding and unfolding. Biochim Biophys Acta Protein Struct Mol Enzymol. 1997;1338(2):161–85.CrossRefGoogle Scholar
  25. 25.
    Pikal MJ, Shah S. The collapse temperature in freeze drying: dependence on measurement methodology and rate of water removal from the glassy phase. Int J Pharm. 1990;62(2):165–86.CrossRefGoogle Scholar
  26. 26.
    Overcashier DE, Patapoff TW, Hsu CC. Lyophilization of protein formulations in vials: investigation of the relationship between resistance to vapor flow during primary drying and small-scale product collapse. J Pharm Sci. 1999;88(7):688–95.CrossRefPubMedGoogle Scholar
  27. 27.
    Depaz RA, Pansare S, Patel SM. Freeze-drying above the glass transition temperature in amorphous protein formulations while maintaining product quality and improving process efficiency. J Pharm Sci. 2016;105(1):40–9.CrossRefPubMedGoogle Scholar
  28. 28.
    Searles J. Observation and implications of sonic water vapor flow during freeze-drying. Am Pharm Rev. 2004;7:58–69.Google Scholar
  29. 29.
    Patel SM, Pikal MJ. Emerging freeze-drying process development and scale-up issues. AAPS PharmSciTech. 2011;12(1):372–8.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Schersch K, Betz O, Garidel P, Muehlau S, Bassarab S, Winter G. Systematic investigation of the effect of lyophilizate collapse on pharmaceutically relevant proteins III: collapse during storage at elevated temperatures. Eur J Pharm Biopharm: Off J Arbeitsgemeinschaft Pharm Verfahrenstechnik. 2013;85(2):240–52.CrossRefGoogle Scholar
  31. 31.
    Carrasquillo KG, Sanchez C, Griebenow K. Relationship between conformational stability and lyophilization-induced structural changes in chymotrypsin. Biotechnol Appl Biochem. 2000;31(1):41–53.CrossRefPubMedGoogle Scholar
  32. 32.
    Griebenow K, Klibanov AM. Lyophilization-induced reversible changes in the secondary structure of proteins. Proc Natl Acad Sci U S A. 1995;92(24):10969–76.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Schneid SC, Startzel PM, Lettner P, Gieseler H. Robustness testing in pharmaceutical freeze-drying: inter-relation of process conditions and product quality attributes studied for a vaccine formulation. Pharm Dev Technol. 2011;16(6):583–90.CrossRefPubMedGoogle Scholar
  34. 34.
    Parker A, Rigby-Singleton S, Perkins M, Bates D, Le Roux D, Roberts CJ, et al. Determination of the influence of primary drying rates on the microscale structural attributes and physicochemical properties of protein containing lyophilized products. J Pharm Sci. 2010;99(11):4616–29.CrossRefPubMedGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2017

Authors and Affiliations

  • Maxwell Korang-Yeboah
    • 1
    Email author
  • Charudharshini Srinivasan
    • 1
  • Akhtar Siddiqui
    • 1
  • David Awotwe-Otoo
    • 2
  • Celia N. Cruz
    • 1
  • Ashraf Muhammad
    • 1
    Email author
  1. 1.Division of Product Quality Research, Office of Testing and Research, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, Food and Drug AdministrationSilver SpringUSA
  2. 2.Division of Post-Marketing Activities II, Office of Lifecycle Drug Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, Food and Drug AdministrationSilver SpringUSA

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