DEHP Nanodroplets Leached From Polyvinyl Chloride IV Bags Promote Aggregation of IVIG and Activate Complement in Human Serum

Abstract

Concerns regarding the impact of subvisible particulate impurities on the safety and efficacy of therapeutic protein products have led manufacturers to implement strategies to minimize protein aggregation and particle formation during manufacturing, storage, and shipping. However, once these products are released, manufacturers have limited control over product handling. In this work, we investigated the effect of di(2-ethylhexyl) phthalate (DEHP) nanodroplets generated in polyvinyl chloride (PVC) bags of intravenous (IV) saline on the stability and immunogenicity of IV immunoglobulin (IVIG) formulations. We showed that PVC IV bags containing saline can release DEHP droplets into the solution when agitated or transported using a pneumatic tube transportation system in a clinical setting. We next investigated the effects of emulsified DEHP nanodroplets on IVIG stability and immunogenicity. IVIG adsorbed strongly to DEHP nanodroplets, forming a monolayer. In addition, DEHP nanodroplets accelerated IVIG aggregation in agitated samples. The immunogenicity of DEHP nanodroplets and IVIG aggregates generated in these formulations were evaluated using an in vitro assay of complement activation in human serum. The results suggested DEHP nanodroplets shed from PVC IV bags could reduce protein stability and induce activation of the complement system, potentially contributing to adverse immune responses during the administration of therapeutic proteins.

Machine Learning and Accelerated Stress Approaches to Differentiate Potential Causes of Aggregation in Polyclonal Antibody Formulations During Shipping

“…The formation of protein aggregates may not only reduce efficacy of therapeutic proteins, but may also provoke adverse immune responses. For example, in vitro studies of IVIg aggregates conducted in human serum show that these aggregates may induce complement activation 11,22 which is associated with adverse events such as anaphylactic shock, altered antibody responses, and impaired immune response. 23,24 Thus, the presence of aggregates in all of our shipped samples at levels that surpassed USP h788i guidelines is concerning.…”

Subvisible Particles in Solutions of Remicade in Intravenous Saline Activate Immune System Pathways in In Vitro Human Cell Systems

“…These droplets have been documented to cause protein aggregation, as well as activation of human complement. 53 Such product degradation due mechanical shock could contribute to adverse immunogenicity and infusion reactions in patients.…”

Effects of Transportation of IV Bags Containing Protein Formulations Via Hospital Pneumatic Tube System: Particle Characterization by Multiple Methods

“…al. that DEHP plasticizer from PVC IV bag material can promote protein aggregation 96 Microparticle levels measured with FlowCam8000 in saline and dextrose from PO IV bags were somewhat higher for bags transported in the PTS than for control bags (Figure 2A-C). Because of the variability observed from bag-to-bag, results are shown for solutions from each individual IV bag.…”

“…The morphologies observed in the particle images were essentially all circular, consistent with droplets of the di(2-ethylhexyl) phthalate (DEHP) plasticizer from the PVC bag material. It has previously been shown by Snell et al 96 that DEHP droplets are dislodged from the bag PVC materials by transportation in a hospital PTS system. This process is due to the severe mechanical shock occurring when the container with the IV bag strikes the wall of system receptacle.…”