ABSTRACT
Purpose
To investigate interactions between protein and silicone oil so that we can provide some mechanistic understanding of protein aggregation in silicone oil lubricated syringes and its prevention by formulation additives such as Polysorbate 80 and Poloxamer 188.
Methods
Interfacial tension values of silicone oil/water interface of abatacept solutions with and without formulation additives were obtained under equilibrium conditions using Attension Theta optical tensiometer. Their adsorption and desorption profiles were measured using Quartz Crystal Microbalancing with Dissipation monitoring (QCM-D). The degree of aggregation of abatacept was assessed based on size exclusion measurement.
Results
Adsorption of abatacept at the oil/water interface was shown. Polysorbat 80 was more effective than Poloxamer 188 in preventing abatacept adsorption. Moreover, it was noted that some of the adsorbed abatacept molecules were not desorbed readily upon buffer rinse. Finally, no homogeneous aggregation was observed at room temperature and a slight increase of aggregation was only observed for samples measured at 40°C which can be prevented using Polysorbate 80.
Conclusions
Interfacial adsorption of proteins is the key step and maybe responsible for the phenomenon of soluble-protein loss when contacting silicone oil and the irreversible adsorption of protein may be associated with protein denaturation/aggregation.
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REFERENCES
Thompson I. New-generation auto-injectors: completing the scale of convenience for self-injection. Drug Delivery Report. 2005; Autum/winter, 47–49.
Jones LS, Kaufmann A, Middaugh CR. Silicone oil induced aggregation of proteins. J Pharm Sci. 2005;94:918–27.
Thirumangalathu R, Krishnan S, Ricci MS, Brems DN, Randolph TW, Carpenter JF. Silicone oil-and agitation-induced aggregation of a monoclonal antibody in aqueous solution. J Pharm Sci. 2009;98:3167–81.
Ludwig DB, Carpenter JF, Hamel JB, Randolph TW. Protein adsorption and excipient effect on kinetic stability of silicone oil emulsions. J Pharm Sci. 2010;99:1721–33.
Ludwig DB, Trotter JT, Gabrielson JP, Carpenter JF, Randolph TW. Flow cytometry: a promising technique for the study of silicone oil-induced particulate formation in protein formulations. Anal Biochem. 2011;410:191–9.
2009. Orencia [product monograph]. Canada: Bristol-Myers Squibb Company.
Rowe RC, Sheskey PJ, Weller PJ. Handbook of pharmaceutical excipients. London: Pharmaceutical; 2003.
Chen P, Prokop RM, Susnar SS, Neumann AW. Interfacial tensions of protein solutions using axisymmetric drop shape analysis. In: Möbius D, Miller R, editors. Proteins at liquid linterfaces. Amsterdam: Elsevier; 1998. p. 303–40.
Messina GML, Satriano C, Marletta G. A multitechnique study of preferential protein adsorption on hydrophobic and hydrophilic plasma-modified polymer surfaces. Colloids Surf B. 2009;70:76–83.
Miller R, Makievski AV, Fainerman VB. Dynamics of adsorption from solutions. In: Fainerman VB, Mobius D, Miller R, editors. Surfactants: chemistry, interfacial properties, applications. Amsterdam: Elsevier, 2001. p. 287–399.
Alexandridis P, Holzwarth JF, Hatton TA. Micellization of poly (ethylene-oxide)-poly (propylene-oxide)-poly (ethylene-oxide) triblock copolymers in aqueous solutions: thermodynamics of copolymer association. Macromolecules. 1994;27:2414–25.
Fainerman VB, Lucassen-Reynders EH, Miller R. Adsorption of surfactants and proteins at fluid interfaces. Colloids and Surf A. 1998;143:141–65.
Albet-Torres N, Gunnarsson A, Persson M, Balaz M, Hook F, Mansson A. Molecular motors on lipid bilayers and silicon dioxide: different driving forces for adsorption. Soft Mater. 2010;6:3211–9.
Berglin M, Pinori E, Sellborn A, Andersson M, Hulander M, Elwing H. Fibrinogen adsorption and conformational change on model polymers: novel aspects of mutual molecular rearrangement. Langmuir. 2009;25:5602–8.
Sauerbrey G. The use of quartz oscillators for weighing thin layers and for microweighing. Zeitschrift fuer Physik. 1959;155:206–22.
Vogt BD, Lin EK, Wu W-L, White CC. Effect of film thickness on the validity of the sauerbrey equation for hydrated polyelectrolyte films. J Phys Chem B. 2004;108:12685–90.
Jordan JL, Fernandez EJ. QCM-D sensitivity to protein adsorption reversibility. Biotechnol Bioeng. 2008;101:837–42.
Tadros TF. Applied surfactants: principles and applications. Weinheim: Wiley-VCH; 2008. p. 100.
Lucassen-Reynders EH. Competitive adsorption of emulsifiers 1. theory for adsorption of small and large molecules. Colloids Surf A. 1994;91:79–88.
Fainerman VB, Lucassen-Reynders EH, Miller R. Description of the adsorption behavior of proteins at water/fluid interfaces in the framework of a two-dimensional solution model. Adv Colloid Interface Sci. 2003;106:237–59.
Fainerman VB, Miller R. Thermodynamics of adsorption of surfactants at the fluid interfaces. In: Fainerman VB, Möbius D, Miller R, editors. Surfactants: chemistry, interfacial properties, applications. Amsterdam: Elsevier; 2001. p. 99–188.
AcknowledgmentS & DISCLOSURES
Authors would like to thank the support of Drug Product Science and Technology Management of Bristol-Myers Squibb Company and Dr. M. Hussain for critical review.
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Appendix
Appendix
Some Theoretical Considerations
In this study, it is assumed that the oil/water (buffer) has a sharp interface (19,20) and its interfacial tension value is γ0 which can be measured, and the adsorption of protein molecules or surfactant molecules or both at the oil/water interface can reduce the interfacial tension to γ. Therefore, the surface pressure is expressed as: Π = γ0-γ, and Π can be modeled as the concentration (Г) in the interfacial layer, which is also a function of protein bulk concentration c, and the interfacial molar area (ω). For a surfactant (component 2) in a protein solution (component 1), the following equation can be derived based on the equation of state assuming the protein is in the state with minimal molar area and the surfactant is in a single adsorption state:
while the expressions for the adsorption isotherm of protein and surfactant are
with
The average molar area of adsorbed component 1 and 2 is
Here c1, c2, ω1, ω2, b1, b2 are the concentrations, molar interfacial area, and bulk/interface distribution coefficients of protein and surfactant, and Γ1 and Γ2 are the concentration of protein and surfactant in the interfacial layer. ael is a parameter related to the electrostatic interaction in the solution depending on the dielectric constant of the protein solution, the total concentration of electrolytes, the number of non-bound unit charges in the protein molecules, etc (20,21). Dynamically, the time-dependent adsorption, Γ(t), depends on the diffusion coefficient of the molecules, bulk concentration and time, and adsorption kinetic model (10) is shown as the following:
where D is the diffusion coefficient and c0 is the bulk concentration, t is the time. In the following text, Eqs. 1–5 will be used as qualitative guidance for discussion. For a simplied system, the above equation can be reduced to
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Li, J., Pinnamaneni, S., Quan, Y. et al. Mechanistic Understanding of Protein-Silicone Oil Interactions. Pharm Res 29, 1689–1697 (2012). https://doi.org/10.1007/s11095-012-0696-6
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DOI: https://doi.org/10.1007/s11095-012-0696-6