Long‐Term Storage of Proteins

John. F. Carpenter1, Mark C. Manning1, Theodore W. Randolph2

1 university of Colorado Health Sciences Center, Denver, 2 University of Colorado, Boulder
Publication Name:  Current Protocols in Protein Science
Unit Number:  Unit 4.6
DOI:  10.1002/0471140864.ps0406s27
Online Posting Date:  May, 2002
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Abstract

This unit provides a summary of some of the issues that researchers face when attempting to store purified proteins. It briefly explains the stresses that induce protein aggregation the major causes for chemical degradation. It also discusses how to use various storage strategies to increase the longā€term stability of proteins. When appropriate it points out critical mistakes to avoid. This unit provides a summary of some of the issues that researchers face when attempting to store purified proteins.

     
 
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Table of Contents

  • Protein Aggregation
  • Chemical Degradation
  • Storage in Nonfrozen Aqueous Solutions
  • Storage as Salted‐Out Precipitates
  • Storage as Frozen Solutions
  • Storage as Freeze‐Dried Solids
  • Selecting an Appropriate Storage Method
  • Inhibition of Proteases
  • Literature Cited
     
 
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Materials

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Figures

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Literature Cited

Literature Cited
   Anchordoquy, T.J. and Carpenter, J.F. 1996. Polymers protect lactate dehydrogenase during freeze‐drying by inhibiting dissociation in the frozen state. Arch. Biochem. Biophys. 332:231‐238.
   Carpenter, J.F. and Chang, B.S. 1996. Lyophilization of protein pharmaceuticals. In Biotechnology and Biopharmaceutical Manufacturing, Processing and Preservation (K. Avis and V. Wu, eds.) pp. 199‐263. Intepharm Press, Buffalo Grove, Ill.
   Carpenter, J.F., Pikal, M.J., Chang, B.S., and Randolph, T.W. 1997. Rational design of stable lyophilized protein formulations: Some practical advice. Pharm. Res. 14:969‐975.
   Chang, B.S., Kendrick, B.S., and Carpenter, J.F. 1996. Surface‐induced denaturation of proteins during freezing and its inhibition by surfactants. J. Pharm. Sci. 85:1325‐1330.
   Cleland, J.L., Powell, M.F., and Shire, S.J. 1993. The development of stable protein formulations: A close look at protein aggregation, deamidation, and oxidation. Crit. Rev. Ther. Drug Carrier Syst. 10:307‐377.
   Costantino, H.R., Langer, R., and Klibanov, A.M. 1994. Solid‐phase aggregation of proteins under pharmaceutically relevant conditions. J. Pharm. Sci. 83:1662‐1669.
   DePaz, R.A., Barnett, C.C., Dale, D.A., Carpenter, J.F., Gaertner, A.L., and Randolph, T.W. 2000. The excluding effects of sucrose on a protein chemical degradation pathway: Methionine oxidation in subtilisin. Arch. Biochem. Biophys. 384:123‐132.
   Heller, M., Carpenter, J.F., and Randolph, T.W. 1996. Effect of phase separating systems on lyophilized hemoglobin. J. Pharm. Sci. 85:1358‐1362.
   Hovorka, S.W., Hong, J., Cleland, J.L., and Schoneich, C. 2001. Metal‐catalyzed oxidation of human growth hormone: Modulation by solvent‐induced changes of protein conformation. J. Pharm. Sci. 90:58‐69.
   Jaenicke, R. 2000. Stability and stabilization of globular proteins in solution. J. Biotech. 79:193‐203.
   Kendrick, B.S., Carpenter, J.F., Cleland, J.L., and Randolph, T.W. 1998. A transient expansion of the native state precedes aggregation of recombinant human interferon‐gamma. Proc. Natl. Acad. Sci. U.S.A. 95:14142‐14146.
   Kim, Y.‐S., Wall, J.S., Meyer, J., Murphy, C.C., Randolph, T.W., Manning, M.C., Solomon, A., and Carpenter, J.F. 2000. Thermodynamic modulation of light chain amyloid fibril formation. J. Biol. Chem. 275:1570‐1574.
   Kosky, A.A., Razzaq, U.O., Treuheit, M.J., and Brems, D.N. 1999. The effects of alpha‐helix on the stability of Asn residues: Deamidation rates in peptides of varying helicity. Protein Sci. 8:2519‐2523.
   Kossiakoff, A.A. 1998. Tertiary structure is a principal determinant to protein deamidation. Science 240:191‐194.
   Manning, M.C., Patel, K., and Borchardt, R.T. 1989. Stability of protein pharmaceuticals. Pharm. Res. 6:903‐918.
   Patel, K. and Borchardt, R.T. 1990a. Chemical pathways of peptide degradation. III. Effect of primary sequence on the pathways of deamidation of asparaginyl residues in hexapeptides. Pharm. Res. 7:787‐793.
   Patel, K. and Borchardt, R.T. 1990b. Chemical pathways of peptide degradation. II. Kinetics of deamidation of an asparaginyl residue in a model hexapeptide. Pharm. Res. 7:703‐711.
   Senderoff, R.I., Kontor, K.M., Kreilgaard, L., Chang, J.J., Patel, S., Krakover, J., Heffernan, J.K., Snell, L.B., and Rosenberg, G.B. 1998. Consideration of conformational transitions and racemization during process development of recombinant glucagon‐like peptide‐1. J. Pharm. Sci. 87:183‐189.
   Timasheff, S.N. 1998. Control of protein stability and reactions by weakly interacting cosolvents: The simplicity of the complicated. Adv. Prot. Chem. 51:355‐432.
   Volkin, D.B., Mach, H., and Middaugh, C.R. 1997. Degradative covalent reactions important to protein stability. Mol. Biotechnol. 8:105‐122.
   Webb, J.N., Webb, S.D., Cleland, J.L., Carpenter, J.F., and Randolph, T.W. 2001. Partial molar volume, surface area, and hydration changes for equilibrium unfolding and formation of aggregation transition state: High‐pressure and cosolute studies on recombinant human IFN‐γ. Proc. Natl. Acad. Sci. U.S.A. 98:7259‐7264.
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