Ultrasensitive Bead-Based Immunoassay for Real-Time Continuous Sample Flow Analysis
The performance of heterophase immunoassays is often limited by the kinetics of analyte binding. This problem is partially solved by bead-based assays, which are characterized by rapid diffusion in the particle suspension. However, at low analyte concentrations, the binding rate is still low. Here,...
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MDPI AG
2025-05-01
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| Series: | Biosensors |
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| author | Yuri M. Shlyapnikov Elena A. Shlyapnikova |
| author_facet | Yuri M. Shlyapnikov Elena A. Shlyapnikova |
| author_sort | Yuri M. Shlyapnikov |
| collection | DOAJ |
| description | The performance of heterophase immunoassays is often limited by the kinetics of analyte binding. This problem is partially solved by bead-based assays, which are characterized by rapid diffusion in the particle suspension. However, at low analyte concentrations, the binding rate is still low. Here, we demonstrate a further improvement of analyte binding kinetics in bead-based immunoassays by simultaneously concentrating both an analyte and magnetic beads in a compact spatial region where binding occurs. The analyte is electrophoretically concentrated in a flow cell where beads are magnetically retained and dragged along the channel by viscous force. The flow cell is integrated with a microarray-based signal detection module, where beads with bound analyte scan the microarray surface and are retained on it by single specific interactions, assuring ultra-high sensitivity of the method. Thus, a continuous flow assay system is formed. Its performance is demonstrated by simultaneous detection of model pathogen biomarkers, cholera toxin (CT) and staphylococcal enterotoxin B (SEB), with a detection limit of 0.1 fM and response time of under 10 min. The assay is capable of real-time online sample monitoring, as shown by a 12 h long continuous flow analysis of tap water for SEB and CT. |
| format | Article |
| id | doaj-art-7adcc77a99ed4d4b970fbf7a67e73598 |
| institution | OA Journals |
| issn | 2079-6374 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
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| series | Biosensors |
| spelling | doaj-art-7adcc77a99ed4d4b970fbf7a67e735982025-08-20T01:56:17ZengMDPI AGBiosensors2079-63742025-05-0115531610.3390/bios15050316Ultrasensitive Bead-Based Immunoassay for Real-Time Continuous Sample Flow AnalysisYuri M. Shlyapnikov0Elena A. Shlyapnikova1Institute of Theoretical and Experimental Biophysics of the Russian Academy of Sciences, Institutskaya 3, 142290 Pushchino, RussiaInstitute of Theoretical and Experimental Biophysics of the Russian Academy of Sciences, Institutskaya 3, 142290 Pushchino, RussiaThe performance of heterophase immunoassays is often limited by the kinetics of analyte binding. This problem is partially solved by bead-based assays, which are characterized by rapid diffusion in the particle suspension. However, at low analyte concentrations, the binding rate is still low. Here, we demonstrate a further improvement of analyte binding kinetics in bead-based immunoassays by simultaneously concentrating both an analyte and magnetic beads in a compact spatial region where binding occurs. The analyte is electrophoretically concentrated in a flow cell where beads are magnetically retained and dragged along the channel by viscous force. The flow cell is integrated with a microarray-based signal detection module, where beads with bound analyte scan the microarray surface and are retained on it by single specific interactions, assuring ultra-high sensitivity of the method. Thus, a continuous flow assay system is formed. Its performance is demonstrated by simultaneous detection of model pathogen biomarkers, cholera toxin (CT) and staphylococcal enterotoxin B (SEB), with a detection limit of 0.1 fM and response time of under 10 min. The assay is capable of real-time online sample monitoring, as shown by a 12 h long continuous flow analysis of tap water for SEB and CT.https://www.mdpi.com/2079-6374/15/5/316bead-based ultrasensitive immunoassaymagnetic beadscholera toxinstaphylococcal enterotoxin B |
| spellingShingle | Yuri M. Shlyapnikov Elena A. Shlyapnikova Ultrasensitive Bead-Based Immunoassay for Real-Time Continuous Sample Flow Analysis Biosensors bead-based ultrasensitive immunoassay magnetic beads cholera toxin staphylococcal enterotoxin B |
| title | Ultrasensitive Bead-Based Immunoassay for Real-Time Continuous Sample Flow Analysis |
| title_full | Ultrasensitive Bead-Based Immunoassay for Real-Time Continuous Sample Flow Analysis |
| title_fullStr | Ultrasensitive Bead-Based Immunoassay for Real-Time Continuous Sample Flow Analysis |
| title_full_unstemmed | Ultrasensitive Bead-Based Immunoassay for Real-Time Continuous Sample Flow Analysis |
| title_short | Ultrasensitive Bead-Based Immunoassay for Real-Time Continuous Sample Flow Analysis |
| title_sort | ultrasensitive bead based immunoassay for real time continuous sample flow analysis |
| topic | bead-based ultrasensitive immunoassay magnetic beads cholera toxin staphylococcal enterotoxin B |
| url | https://www.mdpi.com/2079-6374/15/5/316 |
| work_keys_str_mv | AT yurimshlyapnikov ultrasensitivebeadbasedimmunoassayforrealtimecontinuoussampleflowanalysis AT elenaashlyapnikova ultrasensitivebeadbasedimmunoassayforrealtimecontinuoussampleflowanalysis |