Protein-Corona-by-Design in 2D: A Reliable Platform to Decode Bio-Nano Interactions for the Next-Generation Quality-by-Design Nanomedicines.

Kuo‐Ching Mei,Khuloud T Al‐Jamal,Huw D Summers,Robert C Hider,Er Zhen Teoh,Yueting Li,Volker Mailänder,Belén Ballesteros,Justyna Piasecka,Adam Walters,Artur Ghazaryan

doi:10.1002/adma.201802732

Abstract

Hard corona (HC) protein, i.e., the environmental proteins of the biological medium that are bound to a nanosurface, is known to affect the biological fate of a nanomedicine. Due to the size, curvature, and specific surface area (SSA) 3-factor interactions inherited in the traditional 3D nanoparticle, HC-dependent bio–nano interactions are often poorly probed and interpreted. Here, the first HC-by-design case study in 2D is demonstrated that sequentially and linearly changes the HC quantity using functionalized graphene oxide (GO) nanosheets. The HC quantity and HC quality are analyzed using NanoDrop and label-free liquid chromatography–mass spectrometry (LC-MS) followed by principal component analysis (PCA). Cellular responses (uptake and cytotoxicity in J774 cell model) are compared using imaging cytometry and the modified lactate dehydrogenase assays, respectively. Cellular uptake linearly and solely correlates with HC quantity (R2 = 0.99634). The nanotoxicity, analyzed by retrospective design of experiment (DoE), is found to be dependent on the nanomaterial uptake (primary), HC composition (secondary), and nanomaterial exposure dose (tertiary). This unique 2D design eliminates the size–curvature–SSA multifactor interactions and can serve as a reliable screening platform to uncover HC-dependent bio–nano interactions to enable the next-generation quality-by-design (QbD) nanomedicines for better clinical translation.

Highlights

Hard corona (HC) protein, i.e., the environmental proteins of the biological medium that are bound to a nanosurface, is known to affect the biological fate of a nanomedicine
Investigation of HC on 3D nanoparticles is influenced by the surface charge/surface chemistry and the 3-factor interactions between the size, specific surface area (SSA, the surface area/mass ratio), and the curvature, i.e., both curvature and SSA are sizedependent factors for HC formation.[8] 2D nanomaterials, i.e., nanomaterials where the third dimension is almost negligible (≈1 nm per layer), have much greater SSA when compared to 3D nanomaterials
From Equation (1), cell viability was a collective consequence of all three factors, where the factor A (PC-1b score) has the largest effect followed by factor B (PC-2b score, absolute coefficient = 0.23; (0.23/0.36) × 100% = 64%, i.e., the influence of factor B is ≈64% of factor A), and factor C (graphene dose, absolute coefficient = 0.18; (0.18/0.36) × 100% = 50%, i.e., factor C is half as influential as factor A). These results suggest that the overall cell viability was mainly correlated with cellular uptake/HC quantity

Summary

Introduction

Hard corona (HC) protein, i.e., the environmental proteins of the biological medium that are bound to a nanosurface, is known to affect the biological fate of a nanomedicine. The first correlation study was quantitative cellular uptake using J774 cells (mouse monocyte macrophage) incubated with all four GO derivatives at 20 μg mL−1 concentration for 24 h (see “Methods” and Figure S6 in the Supporting Information for methods and sample preparation).

Results

Conclusion