Zeptosens' protein microarrays: A novel high performance microarray platform for low abundance protein analysis

Abstract

Protein microarrays are considered an enabling technology, which will significantly expand the scope of current protein expression and protein interaction analysis. Current technologies, such as two-dimensional gel electrophoresis (2-DE) in combination with mass spectrometry, allowing the identification of biologically relevant proteins, have a high resolving power, but also considerable limitations. As was demonstrated by Gygi et al. (Proc. Nat. Acad. Sci. USA 2000,97, 9390-9395), most spots in 2-DE, observed from whole cell extracts, are from high abundance proteins, whereas low abundance proteins, such as signaling molecules or kinases, are only poorly represented. Protein microarrays are expected to significantly expedite the discovery of new markers and targets of pharmaceutical interest, and to have the potential for high-throughput applications. Key factors to reach this goal are: high read-out sensitivity for quantification also of low abundance proteins, functional analysis of proteins, short assay analysis times, ease of handling and the ability to integrate a variety of different targets and new assays. Zeptosens has developed a revolutionary new bioanalytical system based on the proprietary planar waveguide technology which allows us to perform multiplexed, quantitative biomolecular interaction analysis with highest sensitivity in a microarray format upon utilizing the specific advantages of the evanescent field fluorescence detection. The analytical system, comprising an ultrasensitive fluorescence reader and microarray chips with integrated microfluidics, enables the user to generate a multitude of high fidelity data in applications such as protein expression profiling or investigating protein-protein interactions. In this paper, the important factors for developing high performance protein microarray systems, especially for targeting low abundant messengers of relevant biological information, will be discussed and the performance of the system will be demonstrated in experimental examples.

Potent organo-osmium compound shifts metabolism in epithelial ovarian cancer cells

“…We studied the downstream effects of oxidative stress and ROS production, given that ROS can target many biological components within the cell, including DNA. Zeptosens RPPAs were used to study the cellular levels of key proteins involved in DNA damage repair (DDR) in compound 1-exposed cells versus controls (29). RPPA measures the abundance of total protein levels and phosphorylated proteins using epitope-specific fluorescently tagged antibodies (SI Appendix, Table S3).…”

Protein Expression Profiling of Breast Cancer Cells by Dissociable Antibody Microarray (DAMA) Staining

“…A number of different sources of peptides and proteins have been used for protein microarray manufacture, including synthetic peptides (7,8), recombinant proteins (9 -11), and monoclonal and polyclonal antibodies (12)(13)(14). Microarrays have been utilized to study protein expression profiles (15)(16)(17), protein-protein interactions (18,19), drug analysis (11,20), and the diagnosis of diseases such as cancer, food allergies, and infection by pathogenic viruses and bacteria (21)(22)(23)(24). Most of the microarrays use the capture microarray platform (3)(4)(5).…”

Reverse‐phase protein arrays for application‐orientated cancer research

“…TSA-based signal amplification, rolling circle strategies, or fluorescent detection in the near infrared (NIR) range (Fig. 1A) are most frequently employed to generate a quantitative readout [5,[56][57][58]. Samples obtained from in vitro experimentation consist of only a single cell type.…”