Kinetic Modeling of the Release of Ethylene Oxide from Sterilized Plastic Containers and its Interaction with Monoclonal Antibodies.
Ethylene oxide (ETO) is commonly used to sterilize plastic containers, but the effects of residual amounts left after sterilization on protein therapeutics are still not well understood. Here we focus primarily on the factors that influence concentrations of ETO migrating from ETO-treated plastic containers into aqueous solution. A study was designed to investigate the kinetics of this process at various temperatures, and the kinetic data could be fit with a model based on a combination of Fickean diffusion and first-order chemical reaction (to account for observed hydrolysis of ETO). The diffusion and reaction rate constants thus obtained obey Arrhenius-like temperature dependence. These results indicate that for analytical methods involving extraction into water, measurements of residual ETO in a container must account for the effects of ETO hydrolysis. Further, the effects of salt concentration and pH of the fluid in the container on accumulated ETO levels were explored. Finally, interactions of ETO with anti-streptavidin (AntiSA) Immunoglobulin G1 (IgG1) and IgG2 antibodies were studied, with ETO adducts found on all methionine residues when incubated in solutions spiked with ETO at concentrations that could be reached (based on the kinetic studies) in ETO-treated plastic vials. Overall, the likelihood of observable ETO-protein modifications upon storage in ETO-sterilized containers will depend on a complex interplay of protein properties, formulation details, storage conditions, and amount of residual ETO initially in the container.
Ethylene oxide (ETO) is commonly used to sterilize plastic containers, but the effects of residual amounts left after sterilization on protein therapeutics are still not well understood. Here we describe experiments exploring the factors that influence concentrations of ETO migrating from ETO-treated plastic containers into aqueous solution over time. Additionally, interactions of ETO with model antibodies were studied, with ETO adducts found on all methionine residues when incubated in solutions spiked with ETO at concentrations that could potentially be reached in ETO-treated plastic vials. Overall, the likelihood of observable ETO-protein modifications upon storage in ETO-sterilized containers will depend on a complex interplay of protein properties, formulation details, storage conditions, and amount of residual ETO initially in the container.
Device Technologies, Process Development, Amgen Inc., Thousand Oaks, CA Bryan.yu@amgen.com.
Bryan Lei
BL
Yu
Attribute Sciences, Process Development, Amgen Inc., Thousand Oaks, CA.
Jun
J
Han
Attribute Sciences, Process Development, Amgen Inc., Thousand Oaks, CA.
Matthew
M
Hammond
Attribute Sciences, Process Development, Amgen Inc., Thousand Oaks, CA.|Current affiliation: Kythera Biopharmaceuticals, Westlake, CA.
Xuemei
X
Wang
Attribute Sciences, Process Development, Amgen Inc., Thousand Oaks, CA.
Qingchun
Q
Zhang
Attribute Sciences, Process Development, Amgen Inc., Thousand Oaks, CA.|Current affiliation: Snapdragon Chemistry, Cambridge, MA.
Andrew
A
Clausen
Device Technologies, Process Development, Amgen Inc., Thousand Oaks, CA.
Ronald
R
Forster
Device Technologies, Process Development, Amgen Inc., Thousand Oaks, CA.
Mingda
M
Eu
D000911:Antibodies, Monoclonal / Q000737:chemistry*; D004347:Drug Interactions; D004355:Drug Stability; D004864:Equipment and Supplies / Q000592:standards; D005027:Ethylene Oxide / Q000737:chemistry*; D007700:Kinetics; D008962:Models, Theoretical; D004364:Pharmaceutical Preparations / Q000737:chemistry; D010969:Plastics / Q000737:chemistry*; D013242:Sterilization; D013696:Temperature
D000911:Antibodies, Monoclonal; D004364:Pharmaceutical Preparations; D010969:Plastics; D005027:Ethylene Oxide
Antibody; Chemical modification; Ethylene oxide; Headspace–gas chromatography; Mass spectrometry; Plastic container