title: Tumor necrosis factor interaction with gold nanoparticles.
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 × 10(6) (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.
authors.affiliation: National Institute of Standards and Technology, Material Measurement Laboratory, Gaithersburg, MD 20899-8520, USA.
authors.forename: De-Hao
authors.initials: DH
authors.lastname: Tsai
authors.forename: Sherrie
authors.initials: S
authors.lastname: Elzey
authors.forename: Frank W
authors.initials: FW
authors.lastname: Delrio
authors.forename: Athena M
authors.initials: AM
authors.lastname: Keene
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Tyner
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Clogston
authors.forename: Robert I
authors.initials: RI
authors.lastname: Maccuspie
authors.forename: Suvajyoti
authors.initials: S
authors.lastname: Guha
authors.forename: Michael R
authors.initials: MR
authors.lastname: Zachariah
authors.forename: Vincent A
authors.initials: VA
authors.lastname: Hackley
mesh_terms: D000327:Adsorption; D000906:Antibodies / Q000276:immunology; D000937:Antigen-Antibody Reactions; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry; D013499:Surface Properties; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000906:Antibodies; D014409:Tumor Necrosis Factor-alpha; D011092:Polyethylene Glycols; D006046:Gold