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The development of new assays that specifically detect intermediate or final products of biotechnological processes or multiple analytes in biomedical research is important for diagnostics and biotechnology. This study presents a microcapsule‐based sandwich assay for detection of proteins and nucleic acids using flow cytometry as an optical readout. The main component of the assay are robust chemically cross‐linked microcapsules that are coated with adaptor proteins such as protein A or streptavidin. In the first approach, the ability of detecting the blood cancer biomarker beta‐2 microglobulin in the fM to pM concentration range with the help of protein A‐coated capsules is demonstrated. In the second approach, streptavidin‐coated capsules are used for detection of nucleic acids in the nM concentration range. The developed assay allows rapid quantitative analyte measurement, while providing high sensitivity and selectivity at very small sample quantities. In the future, protein A‐ and streptavidin‐coated microcapsules can be used as universal tools for detection of a broad range of analytes.

BackgroundMarine diatoms have a higher fucoxanthin content in comparison to macroalgae. Fucoxanthin features many potent bioactive properties, particularly anti-obesity properties. Despite the great potential for harvesting larger amounts of fucoxanthin, the impacts of light quality (light source, intensity, and photoperiod) on fucoxanthin production and the essential proteins involved in fucoxanthin biosynthesis in marine diatoms remain unclear.ResultsIn the present study, Cylindrotheca closterium was selected from four different species of diatoms based on its high fucoxanthin content and productivity. Optimal light conditions (light source, intensity, and regime) were determined by a “Design of Experiment” approach (software MODDE Pro 11 was used). The model indicated that an 18/6 light/darkness regime increased fucoxanthin productivity remarkably as opposed to a 12/12 or 24/0 regime. Eventually, blue light-emitting diode light, as an alternative to fluorescent light, at 100 μmol/m2/s and 18/6 light/darkness regime yielded maximum fucoxanthin productivity and minimal energy consumption. The fucoxanthin production of C. closterium under the predicted optimal light conditions was assessed both in bottle and bag photobioreactors (PBRs). The high fucoxanthin content (25.5 mg/g) obtained from bag PBRs demonstrated the feasibility of large-scale production. The proteomes of C. closterium under the most favorable and unfavorable fucoxanthin biosynthesis light/darkness regimes (18/6 and 24/0, respectively) were compared to identify the essential proteins associated with fucoxanthin accumulation by matrix-assisted laser desorption/ionization-time of flight–mass spectrometry. Six proteins that were up-regulated in the 18/6 regime but down-regulated in the 24/0 were identified as important chloroplastic proteins involved in photosynthesis, energy metabolism, and cellular processes.ConclusionsBlue light-emitting diode light at 100 μmol/m2/s and 18/6 light/darkness regime induced maximum fucoxanthin productivity in C. closterium and minimized energy consumption. The high fucoxanthin production in the bag photobioreactor under optimal light conditions demonstrated the possibility of commercialization. Proteomics suggests that fucoxanthin biosynthesis is intimately associated with the photosynthetic efficiency of the diatom, providing another technical and bioengineering outlook on fucoxanthin enhancement.Electronic supplementary materialThe online version of this article (10.1186/s12934-018-0957-0) contains supplementary material, which is available to authorized users.

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