Abstract
The use of metal immunoprobes, defined as recognition molecules (e.g., antibodies) labeled with metal tags, constitutes an interesting strategy for the analysis of proteins in biological samples. Fluorescent and biocompatible metal nanoclusters (MNC) have been recently established as powerful tags for detection by spectrofluorimetry, but also by elemental mass spectrometry (MS). Detection of such immunoprobes by elemental MS allows not only the qualitative analysis of the proteins but also their absolute quantification. However, the deviation associated with the MNCs polydispersity will limit the analytical precision, particularly in those samples where the concentrations of the sought protein are very low (e.g., single cell analysis). In this work the synthesis of size monodisperse gold nanoclusters (AuNCs) is investigated by using different experimental conditions such as reaction time and temperature, solvent, reducing agent, and pH, among others. Characterization of AuNCs was performed by spectrofluorimetry, dynamic light scattering (DLS) and high resolution transmission electron microscopy (HR-TEM) measurements.
Highlights
Strategies based on the use of metal immunoprobes to quantify biomolecules by inductively coupled plasma mass spectrometry (ICP-MS) have been widely investigated in recent years in the bioanalytical field [1]
The first study carried out for the synthesis of monodisperse AuNCs was the evaluation of the reaction time
Experimental results obtained by dynamic light scattering (DLS) measurements showed that longer reaction times increase the dispersion in size of AuNCs: by decreasing the time from 15 h to 4 h, a significant improvement in polydispersity index (PDI) from 0.4 to 0.28 was observed
Summary
Strategies based on the use of metal immunoprobes to quantify biomolecules by inductively coupled plasma mass spectrometry (ICP-MS) have been widely investigated in recent years in the bioanalytical field [1] This type of approaches is described as the specific interaction between the analyte (e.g., protein biomarker) and the recognition biomolecule (e.g., antibody) previously labelled with several metals or a metal nanostructure which are detected by elemental MS [2]. The metal nanostructures proposed up to date are diverse, from complexes with several atoms (e.g., Maxpar® and DOTA) [3] to metallic nanoparticles (NPs) with typical diameters above 10 nm [3] In this context, metal nanoclusters (MNCs) have been recently introduced as an interesting alternative for the analysis of biomolecules by elemental MS. MNCs can be employed for bimodal detection by fluorescence and elemental MS
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