Peptide-Conjugated Gold Nanoprobe: Intrinsic Nanozyme-Linked Immunsorbant Assay of Integrin Expression Level on Cell Membrane

Abstract

Precisely quantifying the membrane protein expression level on cell surfaces is of vital importance for early cancer diagnosis and efficient treatment. We demonstrate that gold nanoparticle bioconjugated by a rationally designed peptide as nanoprobe possesses selective labeling and accurate quantification capacity of integrin GPIIb/IIIa on the human erythroleukemia cell line. Through selective recognition and marking of integrin, two-photon photoluminescence of the nanoprobe is exploited for direct observation of protein spatial distribution on cell membrane. More importantly, utilizing intrinsic enzyme-like catalysis property of the nanoprobe, the expression level of integrin on human erythroleukemia cells can be quantitatively counted in an amplified and reliable colorimetric assay without cell lysis and protein extraction process. In addition, the analysis of the correlation between the gold nanoparticle and the membrane protein via relevant inductively coupled plasma mass spectrometry measurement verifies the reliability of the new analytical method. It is anticipated that this facile and efficient strategy holds a great promise for a rapid, precise, and reliable quantification of interested functional membrane proteins on the cell surface.

Mimicking horseradish peroxidase and oxidase using ruthenium nanomaterials

“…For a typical oxidation reaction, the time-dependent absorbance of the solution of 0.1 mM TMB and 0. (10,25,50,75, 100 mg mL À1 ) were used to show the chromogenic reactions implying oxidaselike activity. The steady-state kinetic assays of 50 mg mL À1 Ru NPs with TMB (0.05, 0.1, 0.2, 0.5 mM) as the substrate were performed at room temperature.…”

“…[6][7][8] As a promising candidate for articial enzymes, nanomaterials with intrinsic enzyme-like activity known as nanozymes ignite intensive research interest due to their ability to replace specic enzymes in enzyme-based applications. [9][10][11][12][13] The design and development of novel nanostructured materials mimicking the catalytic role of natural enzymes has emerged as a promising eld for both fundamental and applied research. Since the discovery by Yan and coworkers that Fe 3 O 4 nanoparticles possess intrinsic peroxidase-like activity and show potential to replace natural peroxidase in bioassays, 14 substantial research is directed toward the development of nanomaterial catalysts that mimic functions of peroxidase, oxidase or other natural enzymes with the advantages of low cost, controlled synthesis, tunable catalytic activities, and high stability even under severe reaction conditions.…”

Size, Shape, and Protein Corona Determine Cellular Uptake and Removal Mechanisms of Gold Nanoparticles

“…In recent decades, gold nanoparticles (Au NPs) have been widely applied in biomedical fields such as drug delivery, phototherapy, diagnosis, and biosensing due to their unique properties. Designing efficient nanocarriers for detection, imaging, and targeted therapeutic applications of Au NPs demand well understanding of the endocytosis, exocytosis, and their specific mechanisms.…”

Design, fabrication, and biomedical applications of bioinspired peptide–inorganic nanomaterial hybrids

“…In another example, Gao et al reported AuNPs conjugated by a rationally designed peptide (H 2 N-CCYKKKKQAGDV-COOH) as nanoprobe for targeting integrin GPIIb/IIIa. 89 In the presence of H 2 O 2 , peptide-AuNPs acted as robust catalyst to catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to ox-TMB (Fig. 9a).…”

“…c. Schematic illustration of peptide-AuNP-mediated cancer cell immunoassay. d. Linear regression associates HEL cell number with Au concentration (red curve) and catalytic colorimetric variation (blue curve) 89. Reproduced from Gao et al © 2015 American Chemical Society.…”