Hybrid Minimal Core Streptavidin–Obelin as a Versatile Reporter for Bioluminescence‐based Bioassay

Bashmakova, Eugenia E.; Krasitskaya, Vasilisa V.; Kudryavtsev, Alexander N.; Grigorenko, G.M.; Frank, Ludmila A.

doi:10.1111/php.12648

Cited by 13 publications

(13 citation statements)

References 14 publications

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“…The sensitivity of the immunoassay using the heterologous bridge combination system devised in this study indicates that similar methods would be useful as highly sensitive immunoassays for other small peptides. In addition, this assay can be more simplified by using luciferase–streptavidin or obelin–streptavidin fusing proteins published recently . There is a critical need to continue establishment of valid and reliable methods to measure oxytocin in plasma and other biological fluids.…”

Section: Resultsmentioning

confidence: 99%

Development and clinical application of a bioluminescence enzyme immunoassay for oxytocin

et al. 2018

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The development of a highly sensitive analytical method for oxytocin could be useful in the diagnosis and treatment of autistic spectrum disorder. We previously developed a colorimetric enzyme immunoassay (EIA) for plasma oxytocin measurement. In this study, we developed a method to measure oxytocin concentrations using a higher sensitivity bioluminescent EIA. Biotinylated oxytocin bridged with five lysine residues was used in a competitive format. The standard curve range for oxytocin was 1.0 to 1000 pg/assay. In addition, there was good correlation between the colorimetric and bioluminescent immunoassays in terms of measured oxytocin concentration (r = 0.9665, n = 48). The bioluminescent EIA for plasma oxytocin was more rapid and provided higher sensitivity than the colorimetric immunoassay, making it suitable for clinical application.

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Section: Resultsmentioning

confidence: 99%

Development and clinical application of a bioluminescence enzyme immunoassay for oxytocin

et al. 2018

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“…The fusion protein streptavidin to aequorin (STA-AQ) [ 85 ] and minimum-sized core streptavidin to obelin (SAV-OL) [ 86 ] were highly purified from the inclusion bodies in Escherichia coli ( E. coli ) cells and applied to a bioluminescent sandwich immunoassay. The obtained hybrids demonstrated several advantages over similar chemical conjugates: the easiest way of preparation; maximum retention of properties of the initial protein; and as a result, the higher sensitivity of the assay.…”

Section: Analytical Application Of Ca 2+ -Regulmentioning

confidence: 99%

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

Krasitskaya

Bashmakova

Frank

2020

IJMS

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: The functioning of bioluminescent systems in most of the known marine organisms is based on the oxidation reaction of the same substrate—coelenterazine (CTZ), catalyzed by luciferase. Despite the diversity in structures and the functioning mechanisms, these enzymes can be united into a common group called CTZ-dependent luciferases. Among these, there are two sharply different types of the system organization—Ca2+-regulated photoproteins and luciferases themselves that function in accordance with the classical enzyme–substrate kinetics. Along with deep and comprehensive fundamental research on these systems, approaches and methods of their practical use as highly sensitive reporters in analytics have been developed. The research aiming at the creation of artificial luciferases and synthetic CTZ analogues with new unique properties has led to the development of new experimental analytical methods based on them. The commercial availability of many ready-to-use assay systems based on CTZ-dependent luciferases is also important when choosing them by first-time-users. The development of analytical methods based on these bioluminescent systems is currently booming. The bioluminescent systems under consideration were successfully applied in various biological research areas, which confirms them to be a powerful analytical tool. In this review, we consider the main directions, results, and achievements in research involving these luciferases.

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“…1 BL, like chemiluminescence, does not require any light source and is characterized by a high signal to noise ratio without the main issues related to fluorescence detection, such as photobleaching and autofluorescence. 2 Several BL proteins, including luciferases from different species and photoproteins such as aequorin and obelin, have been investigated and used as reporter proteins in drug screening assays, to monitor target pathway activation, or to track cell location over time in small laboratory animals. [2][3][4] Cells can be genetically engineered to express a BL reporter under the regulation of constitutive promoters or inducible transcription elements to respond to different physical and biochemical stimuli.…”

Section: Introductionmentioning

confidence: 99%

New synthetic red- and orange-emitting luciferases to upgrade in vitro and 3D cell biosensing

Calabretta,

Gregucci,

Michelini

2023

Analyst

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Hybrid Minimal Core Streptavidin–Obelin as a Versatile Reporter for Bioluminescence‐based Bioassay

Cited by 13 publications

References 14 publications

Development and clinical application of a bioluminescence enzyme immunoassay for oxytocin

Development and clinical application of a bioluminescence enzyme immunoassay for oxytocin

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

New synthetic red- and orange-emitting luciferases to upgrade in vitro and 3D cell biosensing

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