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.
Similar content being viewed by others
REFERENCES
Tsutomu A, Yoshiko K. Protection of bovine serum albumin from aggregation by Tween 80. J Pharm Sci. 2000;89(5):646–51.
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.
Wang W, Wang JY, Wang DQ. Dual effects of Tween-80 on protein stability. Int J Pharm. 2008;2007(347):31–8.
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
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.
Kerwin BA. Polysorbate 20 and 80 used in protein biotherapeutics: structure and degradation pathways. J Pharm Sci. 2008;97:2924–35.
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.
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.
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.
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.
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.
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.
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.
Horowitz PM. Kinetic control of protein folding by detergent micelle, liposomes, and chaperonins, vol. 526. Washington, DC: American Chemical Society; 1993. p. 156–63.
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.
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.
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
Li S, Schoneich C, Borchardt RT. Chemical instability of protein pharmaceuticals: mechanisms of oxidation and strategies for stabilization. Biotechnol Bioeng. 1995;48:490–500.
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.
Kerwin BA, Remmele Jr RL. Protect from light: photodegradation and protein biologics. J Pharm Sci. 2007;96:1468–79. doi:10.1002/jps.20815.
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.
Jaeger J, Sorensen K, Wolff SP. Peroxide accumulation in detergents. J Biochem Biophys Methods. 1994;29:77–81.
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.
Davies MJ, Truscott RJ. Photo-oxidation of proteins and its role in cataractogenesis. J Photochem Photobiol B. 2001;63:114–25.
Grossweiner LI. Photochemistry of proteins: a review. Curr Eye Res. 1984;3:137–44.
Vlasak J, Ionescu R. Heterogeneity of monoclonal antibodies revealed by charge-sensitive methods. Curr Pharm Biotechnol. 2008;9:468–81.
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.
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.
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
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.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Agarkhed, M., O’Dell, C., Hsieh, MC. et al. Effect of Polysorbate 80 Concentration on Thermal and Photostability of a Monoclonal Antibody. AAPS PharmSciTech 14, 1–9 (2013). https://doi.org/10.1208/s12249-012-9878-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1208/s12249-012-9878-0