Tumor necrosis factor interaction with gold nanoparticles

De-Hao Tsai; Sherrie Elzey; Frank W. DelRio; Athena M. Keene; Katherine M. Tyner; Jeffrey D. Clogston; Robert I. MacCuspie; Suvajyoti Guha; Michael R. Zachariah; Vincent A. Hackley

doi:10.1039/C2NR30415E

Tumor necrosis factor interaction with gold nanoparticles†

De-Hao Tsai,^a Sherrie Elzey,^a Frank W. DelRio,^a Athena M. Keene,^b Katherine M. Tyner,^b Jeffrey D. Clogston,^c Robert I. MacCuspie,^a Suvajyoti Guha,^ad Michael R. Zachariah^ad and Vincent A. Hackley*^a

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* Corresponding authors

^a National Institute of Standards and Technology, Material Measurement Laboratory, Gaithersburg, MD 20899-8520, USA
E-mail: vince.hackley@nist.gov

^b Food and Drug Administration, Center for Drug Evaluation and Research, Silver Spring, MD, USA

^c Nanotechnology Characterization Laboratory, Advanced Technology Program, SAIC-Frederick, Inc., NCI-Frederick, Frederick, MD 21702, USA

^d University of Maryland, Departments of Mechanical Engineering and Chemistry, College Park, MD, USA

Abstract

We report on a systematic investigation of molecular conjugation of tumor necrosis factor-α (TNF) protein onto gold nanoparticles (AuNPs) and the subsequent binding behavior to its antibody (anti-TNF). We employ a combination of physical and spectroscopic characterization methods, including electrospray-differential mobility analysis, dynamic light scattering, polyacrylamide gel electrophoresis, attenuated total reflectance-Fourier transform infrared spectroscopy, fluorescence assay, and enzyme-linked immunosorbent assay. The native TNF used in this study exists in the active homotrimer configuration prior to conjugation. After binding to AuNPs, the maximum surface density of TNF is (0.09 ± 0.02) nm⁻² with a binding constant of 3 × 10⁶ (mol L⁻¹)⁻¹. Dodecyl sulfate ions induce desorption of monomeric TNF from the AuNP surface, indicating a relatively weak intermolecular binding within the AuNP-bound TNF trimers. Anti-TNF binds to both TNF-conjugated and citrate-stabilized AuNPs, showing that non-specific binding is significant. Based on the number of anti-TNF molecules adsorbed, a substantially higher binding affinity was observed for the TNF-conjugated surface. The inclusion of thiolated polyethylene glycol (SH–PEG) on the AuNPs inhibits the binding of anti-TNF, and the amount of inhibition is related to the number ratio of surface bound SH–PEG to TNF and the way in which the ligands are introduced. This study highlights the challenges in quantitatively characterizing complex hybrid nanoscale conjugates, and provides insight on TNF–AuNP formation and activity.

Nanoscale

Tumor necrosis factor interaction with gold nanoparticles†

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