A proteome scale study reveals how plastic surfaces and agitation promote protein aggregation

Abstract

Protein aggregation in biotherapeutics can reduce their activity and effectiveness. It may also promote immune reactions responsible for severe adverse effects. The impact of plastic materials on protein destabilization is not totally understood. Here, we propose to deconvolve the effects of material surface, air/liquid interface, and agitation to decipher their respective role in protein destabilization and aggregation. We analyzed the effect of polypropylene, TEFLON, glass and LOBIND surfaces on the stability of purified proteins (bovine serum albumin, hemoglobin and α-synuclein) and on a cell extract composed of 6000 soluble proteins during agitation (P = 0.1–1.2 W/kg). Proteomic analysis revealed that chaperonins, intrinsically disordered proteins and ribosomes were more sensitive to the combined effects of material surfaces and agitation while small metabolic oligomers could be protected in the same conditions. Protein loss observations coupled to Raman microscopy, dynamic light scattering and proteomic allowed us to propose a mechanistic model of protein destabilization by plastics. Our results suggest that protein loss is not primarily due to the nucleation of small aggregates in solution, but to the destabilization of proteins exposed to material surfaces and their subsequent aggregation at the sheared air/liquid interface, an effect that cannot be prevented by using LOBIND tubes. A guidance can be established on how to minimize these adverse effects. Remove one of the components of this combined stress - material, air (even partially), or agitation - and proteins will be preserved.

Response of Osteoblasts on Amine-Based Nanocoatings Correlates with the Amino Group Density

“…Protein aggregation occurred due to the high shear experienced by the substrate and the high concentration of adsorbed proteins due to the high density of amino groups in the Ti-TMS-PEI. Our results are also consistent with the study of Schvartz et al [ 22 ] on the effect of agitation on protein aggregation and the work of Borzova et al [ 23 ] on the thermal denaturation and aggregation of BSA.…”

“…Another hypothesis for the cause of the positive effects of the amino-rich organic layers is the increased adsorption of serum proteins such as vitronectin or fibronectin and the resulting enhanced interaction of cellular integrin receptors with the coated surface [ 19 , 20 ]. Several studies reported that amino-functionalized surfaces also have a high adsorption capacity of bovine serum albumin (BSA) due to the protein’s high stability and varying surface charges under neutral or acidic conditions [ 21 , 22 , 23 ]. These proteins adsorbed on the surface are essential in the subsequent interaction of an implant with cells, and the protein density on the surface dictates how cells would behave, such as how they would spread or migrate on the surface [ 24 , 25 ].…”