Coadsorption of a Monoclonal Antibody and Nonionic Surfactant at the SiO₂/Water Interface

Abstract

ZL and FP made equal contributions AbstractDuring the formulation of therapeutic monoclonal antibodies (mAbs), nonionic surfactants are commonly added to attenuate structural rearrangement caused by adsorption/desorption at interfaces during processing, shipping and storage. We examined the adsorption of a mAb (COE-3) at the SiO 2 /water interface in the presence of pentaethylene glycol monododecyl ether (C 12 E 5 ), polysorbate 80 (PS80-20EO) and a polysorbate 80 analogue with 7 ethoxylates (PS80-7EO). Spectroscopic ellipsometry (SE) was used to follow COE-3 dynamic adsorption and neutron reflection (NR) was used to determine interfacial structure and composition.Neither PS80-20EO nor C 12 E 5 had a notable affinity for COE-3 or the interface under the conditions studied, and thus did not prevent COE-3 adsorption. In contrast, PS80-7EO did coadsorb but did not influence the dynamic process or the equilibrated amount of absorbed COE-3. Near equilibration, COE-3 underwent structural rearrangement and PS80-7EO started to bind the COE-3 interfacial layer and subsequently formed a well-defined surfactant bilayer via self-assembly. The resultant interfacial layer was comprised of an inner mAb layer of some 70 Å thick and an outer surfactant layer of a further 70 Å, with distinct transitional regions across the mAb-surfactant and surfactant-bulk water boundaries. Once formed, such interfacial layers were very robust and worked to prevent further mAb adsorption, desorption and structural rearrangement. Such robust interfacial layers could be anticipated to exist for formulated mAbs stored in type II glass vials; further research is required to understand the behaviour of these layers for siliconized glass syringes.

Interfacial Adsorption of a Monoclonal Antibody and Its Fab and Fc Fragments at the Oil/Water Interface

“…We show that the behaviour of the mAb and its constituent fragments at the oil/water interface is distinct from that reported previously at air/water and SiO 2 /water interfaces. [20][21][22][23] Our work thus paves the way for future studies directly utilizing PDMS oil to measureing the exact interfacial adsorption of the mAb and its constituent fragments and examining the effects of ionic strength, pH, surfactant co-adsorption, etc., which will advance our understanding of interfacial effects during mAb fill-finish processes and injection device selection. The "straight-through" method is used measure to ( ), the attenuation per unit length of the oil.…”

“…This structural feature is consistent with our previous observed COE-3 conformation adsorbed at the air/water 21 and hydrophilic silica/water interfaces. 22 In their QCM-D study of how several mAbs adsorbed to interfaces coated with hydrophobic silanes, Wiseman and Frank also concluded that monolayers of globular mAbs had been formed at the surface. 27…”

Recent Advances in Studying Interfacial Adsorption of Bioengineered Monoclonal Antibodies

“…A study by Kim et al [77] showed that polysorbates were able to inhibit protein adsorption but did not result in a protein/surfactant complex with an overall lower surface activity. To add to this, Li et al [78] have recently shown that PS 80 and C 12 E 5 could not prevent COE-3 adsorption at the SiO 2 /water interface. When a polysorbate 80 analogue with 7 ethoxylates (PS80-7EO) was used, it co-adsorbed but did not affect the equilibrated amount of adsorption from COE-3.…”

Interfacial Assembly Inspired by Marine Mussels and Antifouling Effects of Polypeptoids: A Neutron Reflection Study

“…Protein adsorption onto solid substrates is a widespread phenomenon, and its prevention is critical for proper functioning of medical devices and implants, as well as in bioseparation and other applications involving surfaces prone to contamination. − The adsorption of proteins in these cases is largely of a nonspecific nature, and there is little control over the structure and composition of the adsorbed protein layer. When proteins from blood are adsorbed onto the surfaces of medical implants, the interfacial processes could undermine device integration at an early stage, trigger adverse cellular responses, and compromise the device’s performance and fate.…”

“…Specular NR is capable of quantitative determination of the thickness and uniformity of an interfacial layer with subnanometer sensitivity along the surface normal direction. ,,,− It effectively determines how the volume fraction (ϕ) of the layer varies with the distance perpendicular to the interface. The technique has been widely used in determining the interfacial layer structure of a variety of soft matters such as surfactants, − peptides, ,,,,, proteins, − antibodies, ,− DNAs, , polymers, and lipids. , As an example system, we employed a set of peptoid peptidomimetic polymers (PMPs) that we have previously demonstrated to prevent protein adsorption and cell attachment above certain chain length-dependent critical grafted chain densities. ,, PMPs form an excellent model brush system because of their exact chain lengths controlled by solid phase synthesis. , …”