Antibody microarrays for high-throughput, multianalyte analysis

Jin, Hongjun; Zangar, Richard C.

doi:10.3233/cbm-2009-0140

Cited by 11 publications

(5 citation statements)

References 65 publications

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“…While important for e.g. cancer biomarker validation studies high inter-array and day-to-day variability may occur [26], [27]. We investigated whether proteins extracted from PFPE blocks are generally suitable for ELISA.…”

Section: Discussionmentioning

confidence: 99%

The PAXgene® Tissue System Preserves Phosphoproteins in Human Tissue Specimens and Enables Comprehensive Protein Biomarker Research

et al. 2013

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Precise quantitation of protein biomarkers in clinical tissue specimens is a prerequisite for accurate and effective diagnosis, prognosis, and personalized medicine. Although progress is being made, protein analysis from formalin-fixed and paraffin-embedded tissues is still challenging. In previous reports, we showed that the novel formalin-free tissue preservation technology, the PAXgene Tissue System, allows the extraction of intact and immunoreactive proteins from PAXgene-fixed and paraffin-embedded (PFPE) tissues. In the current study, we focused on the analysis of phosphoproteins and the applicability of two-dimensional gel electrophoresis (2D-PAGE) and enzyme-linked immunosorbent assay (ELISA) to the analysis of a variety of malignant and non-malignant human tissues. Using western blot analysis, we found that phosphoproteins are quantitatively preserved in PFPE tissues, and signal intensities are comparable to that in paired, frozen tissues. Furthermore, proteins extracted from PFPE samples are suitable for 2D-PAGE and can be quantified by ELISA specific for denatured proteins. In summary, the PAXgene Tissue System reliably preserves phosphoproteins in human tissue samples, even after prolonged fixation or stabilization times, and is compatible with methods for protein analysis such as 2D-PAGE and ELISA. We conclude that the PAXgene Tissue System has the potential to serve as a versatile tissue fixative for modern pathology.

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Section: Discussionmentioning

confidence: 99%

The PAXgene® Tissue System Preserves Phosphoproteins in Human Tissue Specimens and Enables Comprehensive Protein Biomarker Research

et al. 2013

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“…Since these initial reports, functional protein microarrays have been used in many areas of biomedical research, including proteomics and biomarker analysis. In addition, printing methods have evolved to include both contact and noncontact (piezoelectric) printing methods, and all conventional immobilization methods have been utilized to produce microarrays (78)(79)(80)(81)(82).…”

Section: Microarraysmentioning

confidence: 99%

Solid-Phase Biological Assays for Drug Discovery

Forsberg

Sicard²,

Brennan

2014

Annual Rev. Anal. Chem.

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In the past 30 years, there has been a significant growth in the use of solid-phase assays in the area of drug discovery, with a range of new assays being used for both soluble and membrane-bound targets. In this review, we provide some basic background to typical drug targets and immobilization protocols used in solid-phase biological assays (SPBAs) for drug discovery, with emphasis on particularly labile biomolecular targets such as kinases and membrane-bound receptors, and highlight some of the more recent approaches for producing protein microarrays, bioaffinity columns, and other devices that are central to small molecule screening by SPBA. We then discuss key applications of such assays to identify drug leads, with an emphasis on the screening of mixtures. We conclude by highlighting specific advantages and potential disadvantages of SPBAs, particularly as they relate to particular assay formats.

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“…We use a piezoelectric, noncontact, GeSiM Nanoplotter to print our slides, as previously described (Zangar et al, 2006;Gonzalez et al, 2008a;Jin and Zangar, 2010).…”

Section: First Day: Manufacture Chipsmentioning

confidence: 99%

High‐Throughput, Multiplexed Analysis of 3‐Nitrotyrosine in Individual Proteins

Jin

Zangar

2012

CP Toxicology

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Reactive nitrogen species (RNS) and reactive oxygen species (ROS) are derived as a result of inflammation and oxidative stress and can result in protein modifications. As such, these protein modifications are used as biomarkers for inflammation and oxidative stress. In addition, modifications in single-tissue-associated proteins released into blood can provide insight into the tissue localization of the inflammation or oxidative stress. We have developed an enzyme-linked immunosorbent assay antibody microarray platform to analyze the levels of 3-nitrotyrosine in specific proteins in a variety of biological samples, including human plasma and sputum. Selective-capture antibodies are used to immunoprecipitate individual proteins from samples onto isolated spots on the microarray chips. Then, a monoclonal antibody for 3-nitrotyrosine is used to detect the amount of 3-nitrotyrosine on each spot. Our studies suggest that this approach can be used to detect trace amounts of 3-nitrotyrosine in human plasma and sputum. In this paper, we describe our antibody microarray protocol for detecting 3-nitrotyrosine in specific proteins.

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Antibody microarrays for high-throughput, multianalyte analysis

Cited by 11 publications

References 65 publications

The PAXgene® Tissue System Preserves Phosphoproteins in Human Tissue Specimens and Enables Comprehensive Protein Biomarker Research

The PAXgene® Tissue System Preserves Phosphoproteins in Human Tissue Specimens and Enables Comprehensive Protein Biomarker Research

Solid-Phase Biological Assays for Drug Discovery

High‐Throughput, Multiplexed Analysis of 3‐Nitrotyrosine in Individual Proteins

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