Multiplex Label Free Characterization of Cancer Cell Lines Using Surface Plasmon Resonance Imaging

Abstract

Rapid multiplex cell surface marker analysis can expedite investigations in which large number of antigens need to be analyzed. Simultaneous analysis of multiple surface antigens at the same level of sensitivity is however limited in the current golden standard analysis method, flow cytometry. In this paper we introduce a surface plasmon resonance imaging (SPRi)-based technique for 44-plex parameter analysis using a single sample, in less than 20 min. We analyzed the expression on cells from five different cancer cell lines by SPRi on a 44-plex antibody array including 4 negative controls and compared the output with flow cytometry. The combined correlation of the markers that showed expression by flow cytometry was 0.76. The results demonstrate as a proof of principle that SPRi can be applied for rapid semi-quantitative multiplex cell surface marker analysis.

Recent Advancements in Aptamer-Based Surface Plasmon Resonance Biosensing Strategies

“…Multiplexed molecular sensing systems aim to detect multiple targets simultaneously from a single sample, thereby reducing the required volume and minimizing the detection time [74][75][76]. There has been common interest in the development of analytical tools for biomolecule detection using SPR sensors.…”

“…Gyurcsányi et al reported that an optimal surface density of the aptamers yielded the best measured affinity, which is largely determined by the size of the target [80]. Multiplexed molecular sensing systems aim to detect multiple targets simultaneously from a single sample, thereby reducing the required volume and minimizing the detection time [74][75][76]. There has been common interest in the development of analytical tools for biomolecule detection using SPR sensors.…”

Trends in SPR Cytometry: Advances in Label-Free Detection of Cell Parameters

“…Adherent cell and platelets show, both in flow and nonflow conditions, a continuous increasing signal [31], while the RBC signal is stable under flow conditions. Nevertheless, data with adherent cells still yields quantitative information, and can be mixed on an SPRi array to simultaneous label-free detection of 44 antigens on cells within 20 min [16]. A higher SPR response can mean that more cells are captured [32] but also gives information about the antigen expression on the cell surface as this correlates with flow cytometry measurements.…”

“…Additionally, it shows the potential for SPRi cytometry to measure the presence or absence of cell surface antigens on red blood cells (RBCs). Alternatively, SPRi cytometry is described for the ratio of the number of various cell membrane antigens [16]. In Section 3, we summarize a novel SPRi strategy that can be used to rank the avidity of ligands to cellular receptors or avidity of antibody-IgG-opsonized cells (red blood cells, RBCs) to IgG-Fc-receptors (FcγR).…”

Surface Plasmon Resonance Sensors for Concentration and Reaction Kinetic Detections

“…These sensors are applied to the detections of the reactions between protein and protein, protein and DNA/RNA, small molecules and proteins/DNA/RNA, proteins/DNA/RNA/ small molecules and cells, and so on [5][6][7][8]. The application fields include chemical and biological analyses, food safety and environmental monitoring, and drug screening [9,10].…”