| 11. |
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| 12. |
- Neumann, Felix, et al.
(författare)
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Isotachophoretically-driven rolling circle amplification unit for nucleic acid detection
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- Nucleic acid amplification tests have revolutionized the biomedical field by offering high sensitivity and specificity. Polymerase chain reaction (PCR) is considered as the gold standard for nucleic acid amplification; however, it requires sophisticated instrumentation for temperature cycling and real-time detection which makes it expensive. Isothermal amplification technologies have been developed to overcome these drawbacks, such as rolling circle amplification (RCA). In this work, we use the RCA and combine it with isotachophoresis (ITP) to increase the sensitivity for fluorescent real-time detection of nucleic acid amplification. For this, we use a top-down approach by first developing a suitable buffer system that supports RCA and ITP, and subsequently show the focusing of differently-sized and concentrated RCA products. Next, we compare our ITP-RCA assay with a commercial instrument for real-time fluorescence monitoring and demonstrate higher sensitivity from our method. Finally, we aim to combine the ligation and amplification step into ITP to simplify the RCA assay into a one-step reaction. The presented combination of RCA with ITP opens up new opportunities by making nucleic acid detection faster and simpler with potential applications for molecular diagnostics of infectious diseases.
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| 13. |
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| 14. |
- Rokhas, Maria Khihon, et al.
(författare)
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CE analysis of single wood cells performing hydrolysis and preconcentration in open microchannels
- 2014
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Ingår i: Electrophoresis. - : Wiley. - 0173-0835 .- 1522-2683. ; 35:2-3, s. 450-457
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Tidskriftsartikel (refereegranskat)abstract
- In the present work, monosaccharides from pulp samples and single wood fibers were analyzed with CE, using indirect detection due to the lack of chromophores on the monosaccharides. The hydrolysis degradation of cellulose and hemicellulose into monosaccharides was performed using TFA, either in bulk scale or in microscale. In the microscale, one single wood fiber was hydrolyzed in an open microchannel manufactured on a silicon microchip with the dimensions 50 μm × 50 μm (length 1 or 3 cm). The low monosaccharide amounts derived from a single fiber implied that a preconcentration step was necessary to increase the detectability. Thus, an electromigration preconcentration of the hydrolyzed samples was performed within the microchannel, which resulted in a significantly enhanced signal intensity of the monosaccharides. In addition to the experimental study, computer simulations were performed regarding the preconcentration step of monosaccharides. The results from these simulations correlated well with the experimental results.
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| 15. |
- van der Burg, D., et al.
(författare)
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Method development for mono- and disaccharides monitoring in cell culture medium by capillary and microchip electrophoresis
- 2022
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Ingår i: Electrophoresis. - : Wiley. - 0173-0835 .- 1522-2683. ; 43:9-10, s. 922-929
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Tidskriftsartikel (refereegranskat)abstract
- The rapidly growing, competitive biopharmaceutical market requires tight bioprocess monitoring. An integrated, automated platform for the routine online/at-line monitoring of key factors in the cell culture medium could greatly improve process monitoring. Mono- and disaccharides, as the main energy and carbon source, are one of these key factors. A CE-LIF method was developed for the analysis of several mono- and disaccharides, considering requirements and restrictions for analysis in an integrated, automated monitoring platform, such as the possibility for miniaturization to microchip electrophoresis. Analysis was performed after fluorescent derivatization with 8-aminopyrene-1,3,6-trisulfonic acid. The derivatisation reaction and the separation BGE were optimized using design of experiments. The developed method is applicable to the complex matrix of cell culture medium and proved transferable to microchip electrophoresis.
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