Sweeping of Adsorbed Therapeutic Protein on Prefillable Syringes Promotes Micron Aggregate Generation

Abstract

This study evaluated how differences in the surface properties of prefillable syringe barrels and in-solution sampling methods affect micron aggregates and protein adsorption levels. Three syringe types (glass barrel with silicone oil coating [GLS/SO+], glass barrel without silicone oil coating [GLS/SO-], and cyclo-olefin polymer [COP] barrel syringes) were tested with 3 therapeutic proteins (adalimumab, etanercept, and infliximab) using 2 sampling methods (aspiration or ejection). After quiescent incubation, solutions sampled by aspiration exhibited no significant change in micron aggregate concentration in any syringes, whereas those sampled by ejection exhibited increased micron aggregates in both GLS syringe types. Micron aggregate concentration in ejected solutions generally increased with increasing density of adsorbed proteins. Notably, COP syringes contained the lowest micron aggregate concentrations, which were independent of the sampling method. Correspondingly, the adsorbed protein density on COP syringes was the lowest at 1-2 mg/m, which was much less compared with that on GLS syringes and was calculated to be equivalent to only 1-2 protein layers, as visually confirmed by high-speed atomic force microscopy. These data indicate that low-adsorption prefillable syringes should be used for therapeutic proteins because protein aggregate concentration in the ejected solution is elevated by increased protein adsorption to the syringe surface.

An Assessment of the Ability of Submicron- and Micron-Size Silicone Oil Droplets in Dropped Prefillable Syringes to Invoke Early- and Late-Stage Immune Responses

“…Studies have shown that the concentration of particles in protein solutions stored in siliconized PFSs increases not only under accelerated stress conditions but even under quiescent storage conditions. 4 Pharmaceutical products experience agitation, a typical mechanical stress, during shipping and handling, and in response to agitation, SO induces the formation of protein particles in the submicron-and micron-size range. [5][6][7] These protein particles, frequently referred to as subvisible particles, have been found to exhibit enhanced immunogenicity.…”

Characterization of Aggregated Antibody-Silicone Oil Complexes: From Perspectives of Morphology, 3D Image, and Fcγ Receptor Activation

“…Numerous studies have been dedicated to clarify the characteristics of aggregated therapeutic protein-SO complexes. 11,13,14,16,18,28,30,31 However, the characteristics of aggregated antibody-SO complexes, including their concentration, population, shapes, 3D images, and FcgRs activation capability have been obscurely acknowledged so far. Herein we analyzed the agitation-induced aggregated antibody-SO complexes using multifaceted techniques such as flow imaging, confocal fluorescence microscopy, and FcgRs cell-based assay.…”

Immunogenicity of Bioproducts: Cellular Models to Evaluate the Impact of Therapeutic Antibody Aggregates

“…Moreover, nanometer, submicron, and micron protein particles have been evidenced in intravenous saline bags (38,39). Furthermore, micron aggregates were detected in ejected solutions from prefillable syringes due to increased antibody adsorption to the syringe surface (40). Interestingly, it was shown that bedside filtration could significantly reduce the quantity of submicron-and micron-sized aggregates before BP injection (41).…”