Yu, Xiaowen; Scott, Daniel; Dikici, Emre; Joel, Smita; Deo, Sapna; Daunert, Sylvia published an article in 2019, the title of the article was Multiplexing cytokine analysis: towards reducing sample volume needs in clinical diagnostics.Formula: C26H21N3O3 And the article contains the following content:
The trend for improved more precise diagnostics and management of disease heavily relies on the measurement of panels of biomarkers in physiol. samples of patients. Ideally, the ultimate goal would be to detect as many clin. relevant biomarkers as possible in a single drop of blood, achieving quick, sensitive, reproducible, and affordable detection in small volume physiol. samples. Bioluminescent (BL) proteins provide many of the desired characteristics required for such labels, including detection at extremely low concentrations, no interference from physiol. fluids leading to excellent detection limits, and compatibility with many miniaturized systems. However, to date the use of BL proteins has been restricted by their limited multiplexing capabilities. BL proteins typically exhibit a single emission profile and decay kinetics making the simultaneous detection of multiple analytes difficult. Recent progresses in this area include the use of two different engineered luminescent proteins to achieve resolved signals via one-dimensional time resolution This approach, however, to date only lead to a dual analyte detection. Herein, we have demonstrated that using a two-dimensional approach that combines both temporal and spatial resolution, we can expand the multiplexing capabilities of bioluminescent proteins. To that end, the photoprotein aequorin (AEQ) has been employed for the simultaneous detection of three sep. analytes in a single well, differentiated through the use of three discrete time/wavelength windows. Through a combination of site-specific mutations and synthetic coelenterazines “semi-synthetic” AEQ variants have been developed with altered emission profiles and decay kinetics. In this study, two AEQ mutant proteins were genetically conjugated to three pro-inflammatory cytokines (tumor necrosis factor alpha, interleukins 6 and 8) resulting in AEQ-labeled cytokines. These fusion proteins were combined with synthetic coelenterazines resulting in proteins with differing emission maxima and half-lives to allow for the simultaneous detection of all three cytokines in a single sample. The validity of the assay was demonstrated in serum by employing human physiol. samples and comparing our results with com. available individual tests for each of the three cytokines. The experimental process involved the reaction of 8-Benzyl-2-(4-hydroxybenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one(cas: 55779-48-1).Formula: C26H21N3O3
The Article related to aequorin cytokine fusion protein coelenterazine blood analysis interleukin tnfalpha, Biochemical Methods: Spectral and Related Methods and other aspects.Formula: C26H21N3O3