BRET measurement on CCD camera-based microtiter plate readers
NanoBRET assays, which utilize bioluminescence energy resonance transfer (BRET), have been widely adopted in drug discovery for measuring both protein-protein interactions and drug target engagement. While the EnVision and other traditional well-scanning plate readers that measure a single well at a...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-10-01
|
| Series: | SLAS Discovery |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2472555222136865 |
| _version_ | 1811243334904053760 |
|---|---|
| author | Kelvin F. Cho Noelle Javier Kaylee Choi |
| author_facet | Kelvin F. Cho Noelle Javier Kaylee Choi |
| author_sort | Kelvin F. Cho |
| collection | DOAJ |
| description | NanoBRET assays, which utilize bioluminescence energy resonance transfer (BRET), have been widely adopted in drug discovery for measuring both protein-protein interactions and drug target engagement. While the EnVision and other traditional well-scanning plate readers that measure a single well at a time are satisfactory for signal detection for smaller experiments, it becomes challenging to scale these assays to applications that require higher throughput. To address this, we explored the adaptation of the ViewLux and FLIPR plate readers for measuring NanoBRET signal. These plate readers utilize charge-coupled device (CCD) cameras for detection, which enable imaging of the entire assay plate simultaneously. We used tool compounds to generate data from each plate reader and found that the image-based plate readers can be used to measure NanoBRET signals with high S/B and Z´, resulting in comparable IC50 values to those obtained from the EnVision, while requiring less time to complete reads. Consequently, utilization of image-based plate readers for NanoBRET measurement may enable applications that require faster reads, such as for high-throughput screening or kinetics studies. |
| first_indexed | 2024-04-12T14:05:38Z |
| format | Article |
| id | doaj.art-945221b2132745f5975c7ef19d3ace16 |
| institution | Directory Open Access Journal |
| issn | 2472-5552 |
| language | English |
| last_indexed | 2024-04-12T14:05:38Z |
| publishDate | 2022-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | SLAS Discovery |
| spelling | doaj.art-945221b2132745f5975c7ef19d3ace162022-12-22T03:30:06ZengElsevierSLAS Discovery2472-55522022-10-01277413417BRET measurement on CCD camera-based microtiter plate readersKelvin F. Cho0Noelle Javier1Kaylee Choi2Lead Discovery & Characterization, Therapeutic Discovery, Amgen Research, South San Francisco, CA, USALead Discovery & Characterization, Therapeutic Discovery, Amgen Research, South San Francisco, CA, USACorresponding author.; Lead Discovery & Characterization, Therapeutic Discovery, Amgen Research, South San Francisco, CA, USANanoBRET assays, which utilize bioluminescence energy resonance transfer (BRET), have been widely adopted in drug discovery for measuring both protein-protein interactions and drug target engagement. While the EnVision and other traditional well-scanning plate readers that measure a single well at a time are satisfactory for signal detection for smaller experiments, it becomes challenging to scale these assays to applications that require higher throughput. To address this, we explored the adaptation of the ViewLux and FLIPR plate readers for measuring NanoBRET signal. These plate readers utilize charge-coupled device (CCD) cameras for detection, which enable imaging of the entire assay plate simultaneously. We used tool compounds to generate data from each plate reader and found that the image-based plate readers can be used to measure NanoBRET signals with high S/B and Z´, resulting in comparable IC50 values to those obtained from the EnVision, while requiring less time to complete reads. Consequently, utilization of image-based plate readers for NanoBRET measurement may enable applications that require faster reads, such as for high-throughput screening or kinetics studies.http://www.sciencedirect.com/science/article/pii/S2472555222136865 |
| spellingShingle | Kelvin F. Cho Noelle Javier Kaylee Choi BRET measurement on CCD camera-based microtiter plate readers SLAS Discovery |
| title | BRET measurement on CCD camera-based microtiter plate readers |
| title_full | BRET measurement on CCD camera-based microtiter plate readers |
| title_fullStr | BRET measurement on CCD camera-based microtiter plate readers |
| title_full_unstemmed | BRET measurement on CCD camera-based microtiter plate readers |
| title_short | BRET measurement on CCD camera-based microtiter plate readers |
| title_sort | bret measurement on ccd camera based microtiter plate readers |
| url | http://www.sciencedirect.com/science/article/pii/S2472555222136865 |
| work_keys_str_mv | AT kelvinfcho bretmeasurementonccdcamerabasedmicrotiterplatereaders AT noellejavier bretmeasurementonccdcamerabasedmicrotiterplatereaders AT kayleechoi bretmeasurementonccdcamerabasedmicrotiterplatereaders |