LA-ICP-MS with microarray chip sampling to determine holoceruloplasmin in serum

Abstract

An analytical method was developed to determine holoceruloplasmin (holo-CP) concentration in serum using laser ablation inductively coupled plasma-mass spectrometry (LA-ICP-MS). Since each holo-CP in serum contains six Cu ions, the concentration was estimated quantitatively by determining the Cu concentration. For small sample consumption, polydimethylsiloxane (PDMS) microarray chips with 150 mm diameter pillars were manufactured. The surface of each pillar was immobilized by a ceruloplasmin (CP) antibody (IgG) for an antigen-antibody reaction to afford high selectivity for the sampling of total CP in the serum matrix. A calibration curve with a linear regression coefficient of 0.992 was obtained by standard addition methods. For demonstration, 126 pL of human serum was loaded on each pillar of the microarray chips and analyzed by LA-ICP-MS in single-shot mode. The determined concentration of holo-CP was compared with that of the total CP estimated by competitive enzyme linked immunosorbent assay (ELISA), from which the concentration of apoceruloplasmin (apo-CP) was deduced. In conclusion, this analytical method for determining holo-CP concentration with high selectivity and small sample volume can be used in the confirmative diagnosis of diseases related to copper deficiency in clinical samples.

A multi-parametric microarray for protein profiling: simultaneous analysis of 8 different cytochromes via differentially element tagged antibodies and laser ablation ICP-MS

“…1 In recent years the rst promising examples where laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is used for the analysis of Western blots and microarrays in combination with metal tags have been described in the literature. [8][9][10][11][12] Usually these tags are bi-functional linkers containing a reactive group for covalent antibody binding and a chelator for metal complex formation. In LA-ICP-MS a sample aerosol is generated by LA and injected into high temperature plasma where the sample is promptly vaporized, atomized and ionized.…”

“…Recently Joo et al developed a LA-ICP-MS based antibody microarray to detect the copper containing protein holoceruloplasmin in serum with a limit of detection of 46.7 ng mL À1 . 12 In this case the antigen could be detected directly via its hetero-element copper.…”

Quantum Dots Labeling Strategy for “Counting and Visualization” of HepG2 Cells

“…In recent years, inductively coupled plasma mass spectrometry (ICP-MS) as an element-specific detector has become a revolutionary technique for quantitative bioanalysis by introducing inorganic element-labeling strategies . The advantages of ICP-MS include low limits of detection (LOD), low matrix effects, wide dynamic ranges, rapid analysis capabilities, high sensitivity, and high spectral resolution for elements and isotopes, which is commendable for detection of cells with appropriate elemental tags. ,, The accuracy of using ICP-MS to count the cells was largely dependent on the element-labeling efficiency, and the reported elemental tags for ICP-MS bioanalysis include naturally occurring elements, − metal chelates, − metal-containing polymers, ,− and metal-containing nanoparticles. − In our previous work, an ICP-MS-based magnetic immunoassay protocol was proposed for the detection of Jurkat T cells using magnetic nanoparticles (MNPs)-anti-CD3 as capture probes, Au NPs-anti-CD2 as reporter probes. The method was demonstrated to be sensitive, selective, and capable of counting tumor cells in the real blood sample with the LOD of 86 cells.…”

Biological elemental analysis: A cute‐meet of microfluidic device to inductively coupled plasma mass spectrometry

“…A micropillar array immobilizing ceruloplasmin antibody in microfluidic platform was designed for the evaluation of holoceruloplasmin concentration in serum. 42 Using a strategy combined with LA-ICP-MS and enzyme linked immunosorbent assay, the total amount of protein in 126 μl serum was assessed. This method enables the clinical diagnosis of copper deficiency or copper poisoning.…”