Multiplex quadruple bioluminescent assay system
Abstract Bioluminescence (BL) is unique cold body radiation of light, generated by luciferin–luciferase reactions and commonly used in various bioassays and molecular imaging. However, most of the peak emissions of BL populate the blue-yellow region and have broad spectral bandwidths and thus superi...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-10-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-20468-1 |
| _version_ | 1811336289478246400 |
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| author | Genta Kamiya Nobuo Kitada Shojiro Maki Sung Bae Kim |
| author_facet | Genta Kamiya Nobuo Kitada Shojiro Maki Sung Bae Kim |
| author_sort | Genta Kamiya |
| collection | DOAJ |
| description | Abstract Bioluminescence (BL) is unique cold body radiation of light, generated by luciferin–luciferase reactions and commonly used in various bioassays and molecular imaging. However, most of the peak emissions of BL populate the blue-yellow region and have broad spectral bandwidths and thus superimpose each other, causing optical cross-leakages in multiplex assays. This study synthesized a new series of coelenterazine (CTZ) analogues, named K-series, that selectively illuminates marine luciferases with unique, blue-shifted spectral properties. The optical property and specificity of the K-series CTZ analogues were characterized by marine luciferases, with K2 and K5 found to specifically luminesce with ALuc- and RLuc-series marine luciferases, respectively. The results confirmed that the luciferase specificity and color variation of the CTZ analogues minimize the cross-leakages of BL signals and enable high-throughput screening of specific ligands in the mixture. The specificity and color variation of the substrates were further tailored to marine luciferases (or single-chain bioluminescent probes) to create a multiplex quadruple assay system with four integrated, single-chain bioluminescent probes, with each probe designed to selectively luminesce only with its specific ligand (first authentication) and a specific CTZ analogue (second authentication). This unique multiplex quadruple bioluminescent assay system is an efficient optical platform for specific and high-throughput imaging of multiple optical markers in bioassays without optical cross-leakages. |
| first_indexed | 2024-04-13T17:37:17Z |
| format | Article |
| id | doaj.art-c1778a06a943456a8108bd866e7951d4 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-13T17:37:17Z |
| publishDate | 2022-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-c1778a06a943456a8108bd866e7951d42022-12-22T02:37:19ZengNature PortfolioScientific Reports2045-23222022-10-0112111210.1038/s41598-022-20468-1Multiplex quadruple bioluminescent assay systemGenta Kamiya0Nobuo Kitada1Shojiro Maki2Sung Bae Kim3Department of Engineering Science, Graduate School of Informatics and Engineering, The University of Electro-CommunicationsDepartment of Engineering Science, Graduate School of Informatics and Engineering, The University of Electro-CommunicationsDepartment of Engineering Science, Graduate School of Informatics and Engineering, The University of Electro-CommunicationsResearch Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST)Abstract Bioluminescence (BL) is unique cold body radiation of light, generated by luciferin–luciferase reactions and commonly used in various bioassays and molecular imaging. However, most of the peak emissions of BL populate the blue-yellow region and have broad spectral bandwidths and thus superimpose each other, causing optical cross-leakages in multiplex assays. This study synthesized a new series of coelenterazine (CTZ) analogues, named K-series, that selectively illuminates marine luciferases with unique, blue-shifted spectral properties. The optical property and specificity of the K-series CTZ analogues were characterized by marine luciferases, with K2 and K5 found to specifically luminesce with ALuc- and RLuc-series marine luciferases, respectively. The results confirmed that the luciferase specificity and color variation of the CTZ analogues minimize the cross-leakages of BL signals and enable high-throughput screening of specific ligands in the mixture. The specificity and color variation of the substrates were further tailored to marine luciferases (or single-chain bioluminescent probes) to create a multiplex quadruple assay system with four integrated, single-chain bioluminescent probes, with each probe designed to selectively luminesce only with its specific ligand (first authentication) and a specific CTZ analogue (second authentication). This unique multiplex quadruple bioluminescent assay system is an efficient optical platform for specific and high-throughput imaging of multiple optical markers in bioassays without optical cross-leakages.https://doi.org/10.1038/s41598-022-20468-1 |
| spellingShingle | Genta Kamiya Nobuo Kitada Shojiro Maki Sung Bae Kim Multiplex quadruple bioluminescent assay system Scientific Reports |
| title | Multiplex quadruple bioluminescent assay system |
| title_full | Multiplex quadruple bioluminescent assay system |
| title_fullStr | Multiplex quadruple bioluminescent assay system |
| title_full_unstemmed | Multiplex quadruple bioluminescent assay system |
| title_short | Multiplex quadruple bioluminescent assay system |
| title_sort | multiplex quadruple bioluminescent assay system |
| url | https://doi.org/10.1038/s41598-022-20468-1 |
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