Magnetic Silica‐Coated Fluorescent Microspheres (MagSiGlow) for Simultaneous Detection of Tumor‐Associated Proteins

• Published in: Angewandte Chemie International Edition Vol. 63; no. 24; pp. e202318870 - n/a
• Main Authors: Halabi, Elias A., Gessner, Isabel, Yang, Katherine S., Kim, Jae‐Jun, Jana, Rupsa, Peterson, Hannah M., Spitzberg, Joshua D., Weissleder, Ralph
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 10-06-2024
• Edition: International ed. in English
• Subjects: Algorithms; Assaying; bioconjugation; Biomarkers; Biosensing Techniques - methods; Biosensors; Bright plating; Detection limits; ELISA; Equipment costs; Fluorescence; Fluorescent Dyes - chemistry; Humans; Image analysis; Image enhancement; Image processing; Localization; Magnetofluorescence; microarray; Microspheres; Multiplexing; Neoplasm Proteins - analysis; Neoplasm Proteins - metabolism; Proteins; Silica; Silicon Dioxide - chemistry; Tumors; Wavelengths; bioconjugation; microarray; Magnetofluorescence; multiplexing; ELISA
• ISSN: 1433-7851; 1521-3773; 1521-3773
• DOI: 10.1002/anie.202318870

Multiplexed bead assays for solution‐phase biosensing often encounter cross‐over reactions during signal amplification steps, leading to unwanted false positive and high background signals. Current solutions involve complex custom‐designed and costly equipment, limiting their application in simple laboratory setup. In this study, we introduce a straightforward protocol to adapt a multiplexed single‐bead assay to standard fluorescence imaging plates, enabling the simultaneous analysis of thousands of reactions per plate. This approach focuses on the design and synthesis of bright fluorescent and magnetic microspheres (MagSiGlow) with multiple fluorescent wavelengths serving as unique detection markers. The imaging‐based, single‐bead assay, combined with a scripted algorithm, allows the detection, segmentation, and co‐localization on average of 7500 microspheres per field of view across five imaging channels in less than one second. We demonstrate the effectiveness of this method with remarkable sensitivity at low protein detection limits (100 pg/mL). This technique showed over 85 % reduction in signal cross‐over to the solution‐based method after the concurrent detection of tumor‐associated protein biomarkers. This approach holds the promise of substantially enhancing high throughput biosensing for multiple targets, seamlessly integrating with rapid image analysis algorithms.
A rapid multiplexed, on‐bead immune assay using highly fluorescent magnetic microspheres (MagSiGlow) loaded in microarrays to achieve simultaneous detection and co‐localization of 7500 microspheres per image. We demonstrate remarkable sensitivity and an over 85 % reduction in signal cross‐over in the simultaneous detection of tumor‐associated protein biomarkers.