AAPS PharmSciTech

, Volume 14, Issue 1, pp 1–9 | Cite as

Effect of Polysorbate 80 Concentration on Thermal and Photostability of a Monoclonal Antibody

  • Meera Agarkhed
  • Courtney O’Dell
  • Ming-Ching Hsieh
  • Jingming Zhang
  • Joel Goldstein
  • Arvind Srivastava
Research Article

Abstract

Polysorbate 80 is widely used in protein formulations to protect protein against agitation-induced aggregation. In this study, we address concerns about residual peroxide present in Polysorbate 80 on protein stability. Residual peroxide may oxidize active pharmaceutical ingredients leading to reduced stability and may ultimately lead to lower potency and efficacy. The effect of Polysorbate 80 concentration on thermal and photostability of monoclonal antibody of the IgG1 subclass (MAb1) was evaluated at Polysorbate 80 concentrations ranging from 0.00% to 1.00% (w/v). MAb1 samples at 5 mg/mL with various Polysorbate 80 concentrations were subjected to accelerated thermal stress by incubation at 25°C, 40°C, and 50°C for a period of 4 weeks and light stress per ICH guideline Q1B, option 1. Our results show that Polysorbate 80 concentration of 1.00% (w/v) adversely affected thermal and photostability of MAb1. This study demonstrates the importance of carefully choosing Polysorbate 80 concentration in protein formulations to prevent destabilizing effect of Polysorbate 80 on thermal and photostability.

Key words

aggregation monoclonal antibody non-ionic surfactant oxidation photostability Polysorbate 80 thermal stability 

Notes

Acknowledgment

The authors would like to thank Stephen Tracy for performing the Polysorbate 80 assay on photostability samples.

Conflict of Interest Declaration

The authors have no personal financial or nonfinancial conflicts of interest in the publication of results contained in this manuscript.

REFERENCES

  1. 1.
    Tsutomu A, Yoshiko K. Protection of bovine serum albumin from aggregation by Tween 80. J Pharm Sci. 2000;89(5):646–51.CrossRefGoogle Scholar
  2. 2.
    Damodaran S, Song KB. Kinetics of adsorption of proteins at interfaces: role of protein conformation in diffusional adsorption. Biochim Biophys Acta. 1988;954:253–64.PubMedCrossRefGoogle Scholar
  3. 3.
    Wang W, Wang JY, Wang DQ. Dual effects of Tween-80 on protein stability. Int J Pharm. 2008;2007(347):31–8.CrossRefGoogle Scholar
  4. 4.
    Mahler H-C, Huber F, Kishore RSK, Reindl J, Ruckert P, Muller R. Adsorption behavior of a surfactant and a monoclonal antibody to sterilizing-grade filters. J Pharm Sci 2010; doi:10.1002/jps.22045
  5. 5.
    Ha E, Wang W, Wang YJ. Peroxide formation in polysorbate 80 and protein stability. J Pharm Sci. 2002;91:2252–64. doi:10.1002/jps.10216.PubMedCrossRefGoogle Scholar
  6. 6.
    Kerwin BA. Polysorbate 20 and 80 used in protein biotherapeutics: structure and degradation pathways. J Pharm Sci. 2008;97:2924–35.PubMedCrossRefGoogle Scholar
  7. 7.
    Jones LS, Bam NB, Randolph TW. Surfactant-stabilized protein formulations: a review of protein-surfactant interactions and novel analytical methodologies. ACS Symp Ser. 1997;676:206–22. Chapter 12.CrossRefGoogle Scholar
  8. 8.
    Randolph TW, Jones LS. Surfactant-protein interactions. Rational design of stable protein formulations. In: Carpenter JF, Manning MC, editors. Pharm Biotechnol, vol. 13. New York: Kluwer Academic/Plenum Publishers; 2002. p. 159–75.Google Scholar
  9. 9.
    Bam NB, Randolph TW, Cleland JL. Stability of protein formulations: investigation of surfactant effects by a novel EPR spectroscopic technique. Pharm Res. 1995;12(1):2–11.PubMedCrossRefGoogle Scholar
  10. 10.
    Bam NB, Cleland JL, Yang J, Manning MC, Carpenter JF, Kelley RF, Randolph TW. Tween protects recombinant human growth hormone against agitation-induced damage via hydrophobic interactions. J Pharm Sci. 1998;12:1554–9.CrossRefGoogle Scholar
  11. 11.
    Chou DK, Krishnamurthy R, Randolph TW, Carpenter JF, Manning MC. Effects of Tween 20 and Tween 80 on the stability of albutropin during agitation. J Pharm Sci. 2005. doi:10.1002/jps.20365.
  12. 12.
    Bam NB, Cleland JL, Randolph TW. Molten globule intermediate of recombinant human growth hormone: stabilization with surfactants. Biotechnol Prog. 1996;12:801–9. doi:10.1021/bp960068b.PubMedCrossRefGoogle Scholar
  13. 13.
    Kreilgaard L, Jones LS, Randolph TW, Frokjaer S, Flink JM, Manning MC, Carpenter JF. Effect of Tween 20 on freeze-thawing- and agitation induced aggregation of recombinant human factor XIII. J Pharm Sci. 1998;87:1597–603.PubMedGoogle Scholar
  14. 14.
    Horowitz PM. Kinetic control of protein folding by detergent micelle, liposomes, and chaperonins, vol. 526. Washington, DC: American Chemical Society; 1993. p. 156–63.Google Scholar
  15. 15.
    Lee HJ, McAuley A, Schilke KF, McGuire J. Molecular origins of surfactant-mediated stabilization of protein drugs. Adv Drug Deliv Rev. 2011;63:1160–71.PubMedCrossRefGoogle Scholar
  16. 16.
    Ding S. Quantitation of hydroperoxides in the aqueous solutions of non-ionic surfactants using Polysorbate 80 as the model surfactant. J Pharm Biomed Anal. 1993;11:95–101.PubMedCrossRefGoogle Scholar
  17. 17.
    Wasylaschuk WR, Harmon PA, Wagner G, Harman AB, Templeton AC, Xu H, Reed RA. Evaluation of hydroperoxides in common pharmaceutical excipients. J Pharm Sci 2006. doi:10.1002/jps.20726
  18. 18.
    Li S, Schoneich C, Borchardt RT. Chemical instability of protein pharmaceuticals: mechanisms of oxidation and strategies for stabilization. Biotechnol Bioeng. 1995;48:490–500.PubMedCrossRefGoogle Scholar
  19. 19.
    Singh SR, O’Dell C, Zhang J, Hsieh M, Goldstein J, Liu J, Srivastava A. Effect of Polysorbate 80 quality on the photostability of a monoclonal antibody. AAPS PharmSciTech. 2012;13(2):422–31.PubMedCrossRefGoogle Scholar
  20. 20.
    Kerwin BA, Remmele Jr RL. Protect from light: photodegradation and protein biologics. J Pharm Sci. 2007;96:1468–79. doi:10.1002/jps.20815.PubMedCrossRefGoogle Scholar
  21. 21.
    Manning MC, Chou DK, Murphy BM, Payne RW, Katayama DS. Stability of protein pharmaceuticals: an update. Pharm Res. 2010;27:544–75. doi:10.1007/s11095-009-0045-6.PubMedCrossRefGoogle Scholar
  22. 22.
    Jaeger J, Sorensen K, Wolff SP. Peroxide accumulation in detergents. J Biochem Biophys Methods. 1994;29:77–81.PubMedCrossRefGoogle Scholar
  23. 23.
    Lam XM, Yang JY, Cleland JL. Antioxidants for prevention of methionine oxidation in recombinant monoclonal antibody HER2. J Pharm Sci. 1997;86:1250–5. doi:10.1021/js970143s.PubMedCrossRefGoogle Scholar
  24. 24.
    Davies MJ, Truscott RJ. Photo-oxidation of proteins and its role in cataractogenesis. J Photochem Photobiol B. 2001;63:114–25.PubMedCrossRefGoogle Scholar
  25. 25.
    Grossweiner LI. Photochemistry of proteins: a review. Curr Eye Res. 1984;3:137–44.PubMedCrossRefGoogle Scholar
  26. 26.
    Vlasak J, Ionescu R. Heterogeneity of monoclonal antibodies revealed by charge-sensitive methods. Curr Pharm Biotechnol. 2008;9:468–81.PubMedCrossRefGoogle Scholar
  27. 27.
    Qi P, Volkin DB, Zhao H, Nedved ML, Hughes R, Bass R, Yi SC, Panek ME, Wang D, Dalmonte P, Bond MD. Characterization of the photodegradation of a human IgG1 monoclonal antibody formulated as a high-concentration liquid dosage form. J Pharm Sci. 2009;98:3117–30. doi:10.1002/jps.21617.PubMedCrossRefGoogle Scholar
  28. 28.
    Mason BD, Schöneich C, Kerwin BA. Effect of pH and light on aggregation and conformation of an IgG1 mAb. Mol Pharm. 2012;9:774–90. doi:10.1021/mp2004719.PubMedCrossRefGoogle Scholar
  29. 29.
    Kishore RSK, Pappenberger A, Dauphin IB, Ross A, Buergi B, Staempfli A, Mahler Ha-C. Degradation of Polysorbates 20 and 80: Studies on thermal autoxidation and hydrolysis. Wiley Online Library. Available from http://wileyonlinelibrary.com. doi:10.1002/jps.22290

Copyright information

© American Association of Pharmaceutical Scientists 2012

Authors and Affiliations

  • Meera Agarkhed
    • 1
  • Courtney O’Dell
    • 1
  • Ming-Ching Hsieh
    • 2
  • Jingming Zhang
    • 2
  • Joel Goldstein
    • 1
  • Arvind Srivastava
    • 1
  1. 1.Formulation Development departmentImClone Systems, A Wholly owned subsidiary of Eli Lilly and CompanyBranchburgUSA
  2. 2.Bio Analytical Sciences departmentImClone Systems, A Wholly owned subsidiary of Eli Lilly and CompanyBranchburgUSA

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