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.
Highlights
The desire for high throughput multiplex cell analysis has grown because diseases like cancer involve complex cell surface antigen expression patterns
The sensorgrams show a great divergence of expression on MCF7 cells and likewise different levels of HER2 expression on the five cell lines are seen
The ABC values were normalized to the surface plasmon resonance imaging (SPRi) values in such a way that all ABC values were divided by the highest ABC value times the highest Surface Plasmon Resonance (SPR) value
Summary
The desire for high throughput multiplex cell analysis has grown because diseases like cancer involve complex cell surface antigen expression patterns. Several techniques like fluorescent microscopy and flow cytometry (Fluorescence-Activated Cell Sorting or “FACS”) have proven to be useful for multiplex analysis, but they are time consuming and multiplexing is limited by the available stains and filters of the respective set up. FACS remains the standard for cellular biomarker analysis, new applications of existing techniques are emerging. The authors describe a technique that cycles fluorescence tagging, imaging and bleaching in situ. The uniqueness of this technique is that it can map hundreds of different proteins in one sample as it visualizes molecular clusters as a so-called toponome map.
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