Design of Freeze-Drying Processes for Pharmaceuticals: Practical Advice | Pharmaceutical Research | Springer Nature Link Skip to main content Advertisement Design of Freeze-Drying Processes for Pharmaceuticals: Practical Advice Published: February 2004 Volume 21 , pages 191–200 ( 2004 ) Cite this article Save article View saved research Pharmaceutical Research Aims and scope Submit manuscript Abstract Design of freeze-drying processes is often approached with a “trial and error” experimental plan or, worse yet, the protocol used in the first laboratory run is adopted without further attempts at optimization. Consequently, commercial freeze-drying processes are often neither robust nor efficient. It is our thesis that design of an “optimized” freeze-drying process is not particularly difficult for most products, as long as some simple rules based on well-accepted scientific principles are followed. It is the purpose of this review to discuss the scientific foundations of the freeze-drying process design and then to consolidate these principles into a set of guidelines for rational process design and optimization. General advice is given concerning common stability issues with proteins, but unusual and difficult stability issues are beyond the scope of this review. Control of ice nucleation and crystallization during the freezing step is discussed, and the impact of freezing on the rest of the process and final product quality is reviewed. Representative freezing protocols are presented. The significance of the collapse temperature and the thermal transition, denoted T g `, are discussed, and procedures for the selection of the “target product temperature” for primary drying are presented. Furthermore, guidelines are given for selection of the optimal shelf temperature and chamber pressure settings required to achieve the target product temperature without thermal and/or mass transfer overload of the freeze dryer. Finally, guidelines and “rules” for optimization of secondary drying and representative secondary drying protocols are presented. This is a preview of subscription content, log in via an institution to check access. Access this article Log in via an institution Subscribe and save Springer+ from €37.37 /Month Starting from 10 chapters or articles per month Access and download chapters and articles from more than 300k books and 2,500 journals Cancel anytime View plans Buy Now Buy article PDF 39,95 € Price includes VAT (China (P.R.)) Instant access to the full article PDF. Institutional subscriptions Similar content being viewed by others Practical Advice on Scientific Design of Freeze-Drying Process: 2023 Update Article Open access 02 October 2023 Recommended Best Practices in Freeze Dryer Equipment Performance Qualification: 2022 Article Open access 26 January 2023 Chapter 21: Rational Design of a Freeze-Drying Process for Protein Products Chapter © 2020 Explore related subjects Discover the latest articles, books and news in related subjects, suggested using machine learning. Design of Experiments Protein Design Process Chemistry Process Engineering Thermal Process Engineering Bioprocess Engineering references M. J. Pikal. Freeze-drying of proteins. Part I: process design. BioPharm 3 :18-28 (1990). Google Scholar M. J. Pikal. Freeze-drying of proteins part II: formulation selection. BioPharm 3 :26-30 (1990). Google Scholar M. J. Pikal. Freeze-Drying of Proteins. In V. H. L. Lee (ed.), Peptide and Proteins Delivery , Marcel Dekker, New York, 1998. Google Scholar J. F. Carpenter and B. S. Chang. Lyophilization of protein pharmaceuticals. In K. Avisand and V. Wu (eds.), Biotechnology and Biopharmaceutical Manufacturing, Processing and Preservation , Intepharm Press, Buffalo Grove, IL, 1996, pp. 199-263. Google Scholar J. M. Beals, M. J. Edwards, M. J. Pikal, and J. V. Rinella, Jr. Formulations of obesity protein, Eur. Pat. Appl. (Eli Lilly and Co., USA). EP, 1997, pp. 48 S. L. Nail and L. A. Gatin. Freeze-drying: principles and practice. In K. E. Avis, H. A. Lieberman, and L. Lechman (eds.), Pharmaceutical Dosage Forms: Parenteral Medications , Vol. 2, Marcel Dekker, New York, 1993, pp. 163-233. Google Scholar F. Franks. Freeze drying: from empiricism to predictability. Cryo-Letters 11 :93-110 (1990). Google Scholar J. F. Carpenter, M. J. Pikal, B. S. Chang, and T. W. Randolph. Rational design of stable lyophilized protein formulations: some practical advice. Pharm. Res. 14 :969-975 (1997). Google Scholar A. P. Mackenzie. Basic principles of freeze-drying for pharmaceuticals. Bull. Parenter. Drug Assoc. 20 :101-130 (1966). Google Scholar M. J. Pikal and S. Shah. The collapse temperature in freeze drying: dependence on measurement methodology and rate of water removal from the glassy phase. Int. J. Pharm. 62 :165-186 (1990). Google Scholar E. Y. Shalaev and F. Franks. Changes in the physical state of model mixtures during freezing and drying: impact on product quality. Cryobio. 33 :14-26 (1996). Google Scholar P. L. Privalov. Cold denaturation of proteins. Crit. Rev. Biochem. Mol. Biol. 25 :281-305 (1990). Google Scholar Y. V. Griko, S. Y. Venyaminov, and P. L. Privalov. Heat and cold denaturation of phosphoglycerate kinase (interaction of domains). FEBS Lett. 244 :276-278 (1989). Google Scholar B. S. Chang, B. S. Kendrick, and J. F. Carpenter. Surface-induced denaturation of proteins during freezing and its inhibition by surfactants. J. Pharm. Sci. 85 :1325-1330 (1996). Google Scholar J. F. Carpenter, S. J. Prestrelski, and T. Arakawa. Separation of freezing-and drying-induced denaturation of lyophilized proteins using stress-specific stabilization. I. Enzyme activity and calorimetric studies. Arch. Biochem. Biophys. 303 :456-464 (1993). Google Scholar M. J. Pikal. Lyophilization. In J. Swarbrick and J. Boylan (eds.), Encyclopedia of Pharmaceutical Technology , Marcel Dekker, New York, 2002, pp. 1299-1326. Google Scholar N. Murase and F. Franks. Salt precipitation during the freeze-concentration of phosphate buffer solutions. Biophys. Chem. 34 :293-300 (1989). Google Scholar B. A. Szkudlarek, T. J. Anchordoquy, G. A. Garcia, M. J. Pikal, J. F. Carpenter, and N. Rodriguez-Hornedo. pH changes of phosphate buffer solutions during freezing and their influence on the stability of a model protein, lactate dehydrogenase, Book of Abstracts, 211th ACS National Meeting , New Orleans, LA, 1996, pp. BIOT-138. E. Shalaev, T. Johnson-Elton, L. Change, and M. J. Pikal. Thermophysical properties of pharmaceutically compatible buffers at sub-zero temperatures: implications for freeze drying. Pharm. Res. 19 :195-211 (2002). Google Scholar S. Jiang and S. L. Nail. Effect of process conditions on recovery of protein activity after freezing and freeze-drying. Eur. J. Pharm. Biopharm. 45 :249-257 (1998). Google Scholar J. A. Searles, J. F. Carpenter, and T. W. Randolph. The ice nucleation temperature determines the primary drying rate of lyophilization for samples frozen on a temperature-controlled shelf. J. Pharm. Sci. 90 :860-871. M. C. Heller, J. F. Carpenter, and T. W. Randolph. Protein formulation and lyophilization cycle design: prevention of damage due to freeze-concentration induced phase separation. Biotechnol. Bioeng. 63 :166-174 (1999). Google Scholar M. C. Heller, J. F. Carpenter, and T. W. Randolph. Application of a thermodynamic model to the prediction of phase separations in freeze-concentrated formulations for protein lyophilization. Arch. Biochem. Biophys. 363 :191-201 (1999). Google Scholar K-I. Izutsu, S. Yoshioka, and S. Kojima. Phase separation and crystallization of components in frozen solutions: effect of molecular compatibility between solutes. ACS Symp. Ser. 675 :109-118 (1997). Google Scholar B. Lueckel, D. Bodmer, B. Helk, and H. Leuenberger. Formulations of sugars with amino acids or mannitol-influence of concentration ratio on the properties of the freeze-concentrate and the lyophilizate. Pharm. Dev. Technol. 3 :325-336 (1998). Google Scholar N. A. Williams. Y. Lee, G. P. Polli, and T. A. Jennings. The effects of cooling rate on solid phase transitions and associated vial breakage occurring in frozen mannitol solutions. J. Parenter. Sci. Technol. 40 :135-141 (1986). Google Scholar A. Pyne, R. Surana, and R. Suryanarayanan. Crystallization of mannitol below Tg′ during freeze-drying in binary and ternary aqueous systems. Pharm. Res. 19 :901-908 (2002). Google Scholar M. J. Pikal, S. Shah, D. Senior, and J. E. Lang. Physical chemistry of freeze-drying: measurement of sublimation rates for frozen aqueous solutions by a microbalance technique. J. Pharm. Sci. 72 :635-650 (1983). Google Scholar J. A. Searles, J. F. Carpenter, and T. W. Randolph. Annealing to optimize the primary drying rate, reduce freezing-induced drying rate heterogeneity, and determine Tg′ in pharmaceutical lyophilization. J. Pharm. Sci. 90 : 872-887. M. J. Pikal, S. Shah, M. L. Roy, and R. Putman. The secondary drying stage of freeze drying: drying kinetics as a function of temperature and chamber pressure. Int. J. Pharm. 60 :203-217 (1990). Google Scholar R. P. Bhattacharyya and T. R. Sosnick. Viscosity dependence of the folding kinetics of a dimeric and monomeric coiled coil. Biochem. 38 :2601-2609 (1999). Google Scholar H. A. Kramers. Brownian motion in a field of force and the diffusion model of chemical reactions. Physica 7 :284-304 (1940). Google Scholar X. Ma, D. Q. Wang, R. Bouffard, and A. MacKenzie. Characterization of murine monoclonal antibody to tumor necrosis factor (TNF-MAb) formulation for freeze-drying cycle development. Pharm. Res. 18 :196-202 (2001). Google Scholar M. J. Pikal. Mechanisms of protein stabilization during freeze-drying and storage: the relative importance of thermodynamic stabilization and glassy state relaxation dynamics. Drugs Pharm. Sci. 96 :161-198 (1999). Google Scholar M. J. Pikal and J. E. Lang. Rubber closures as a source of haze in freeze dried parenterals: test methodology for closure evaluation. J. Parenter. Drug Assoc. 32 :162-173 (1978). Google Scholar M. J. Pikal, M. L. Roy, and S. Shah. Mass and heat transfer in vial freeze-drying of pharmaceuticals: role of the vial. J. Pharm. Sci. 73 :1224-1237 (1984). Google Scholar N. Milton, M. J. Pikal, M. L. Roy, and S. L. Nail. Evaluation of manometric temperature measurement as a method of monitoring product temperature during lyophilization. PDA J. Pharm. Sci. Technol. 51 :7-16 (1997). Google Scholar X. C. Tang, S. L. Nail, and M. J. Pikal. Mass transfer in freeze drying: measurement of dry layer resistance by a non-steady state method (the MTM procedure). 1999 AAPS Anual Meeting , New Orleans, Louisiana, 1999. X. C. Tang, S. L. Nail, and M. J. Pikal. Freeze drying process optimization by manometric temperature measurement, 2001 AAPS Annual Meeting , Denver, Colorado, 2001. M. J. Pikal. 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. 39 :115-139 (1985). Google Scholar M. L. Roy and M. J. Pikal. Process control in freeze drying: determination of the end point of sublimation drying by an electronic moisture sensor. J. Parenter. Sci. Technol. 43 :60-66 (1989). Google Scholar M. J. Pikal, K. M. Dellerman, and M. L. Roy. Formulation and stability of freeze-dried proteins: effects of moisture and oxygen on the stability of freeze-dried formulations of human growth hormone. Develop. Bio. Standard. 74 :21-38 (1991). Google Scholar M. J. Hageman. The role of moisture in protein stability. Drug Dev. Ind. Pharm. 14 :2047-2070 (1988). Google Scholar M. S. Kamat, R. A. Lodder, and P. P. DeLuca. Near-infrared spectroscopic determination of residual moisture in lyophilized sucrose through intact glass vials. Pharm. Res. 6 :961-965 (1989). Google Scholar I. Presser, N. Denkinger, H. Hormann, and G. Winter. New methods in monitering of freeze-drying: near-infrared spectroscopy determination of residue moiture during freeze-drying, 2002 Protein Stability Conference , Breckenridge, Colorado, 2002. E. Maltini. Estimation of the viscosities of the frozen sucrose-water system from glass transition temperatures. Ita. J. Food Sci. 4 :371-377 (1993). Google Scholar Download references Author information Authors and Affiliations School of Pharmacy, University of Connecticut, Storrs, Connecticut, 06269-2092 Xiaolin (Charlie) Tang & Michael J. Pikal Authors Xiaolin (Charlie) Tang View author publications Search author on: PubMed Google Scholar Michael J. Pikal View author publications Search author on: PubMed Google Scholar Corresponding author Correspondence to Michael J. Pikal . Rights and permissions Reprints and permissions About this article Cite this article Tang, X.(., Pikal, M.J. Design of Freeze-Drying Processes for Pharmaceuticals: Practical Advice. Pharm Res 21 , 191–200 (2004). https://doi.org/10.1023/B:PHAM.0000016234.73023.75 Download citation Issue date : February 2004 DOI : https://doi.org/10.1023/B:PHAM.0000016234.73023.75 Share this article Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link is not currently available for this article. Copy shareable link to clipboard Provided by the Springer Nature SharedIt content-sharing initiative lyophilization freeze drying secondary drying formulation Access this article Log in via an institution Subscribe and save Springer+ from €37.37 /Month Starting from 10 chapters or articles per month Access and download chapters and articles from more than 300k books and 2,500 journals Cancel anytime View plans Buy Now Buy article PDF 39,95 € Price includes VAT (China (P.R.)) Instant access to the full article PDF. Institutional subscriptions Advertisement