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Optimizing azide-alkyne cycloaddition reactions in terms of toxicity, reactivity and reaction rates.

Fentsahm, P.J. (2021) Optimizing azide-alkyne cycloaddition reactions in terms of toxicity, reactivity and reaction rates.

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Abstract:In this paper, azide-alkyne cycloaddition on fixed SH-EP2 cells was optimized including optimizations in labeling specificity, reaction time, reaction buffers and toxicity. The optimized reaction composition was then tested on live labeling and cell viability. Despite high labeling yields, the SPAAC reaction showed major disadvantages compared to the tested CuAAC reactions with its significantly longer reaction time and high amounts of non-specific labeling which could not be resolved by treatment with Iodoacetamide (IAM). Therefore, further research in this paper was aimed at improving the CuAAC reaction mechanism. Here a new CuAAC reaction composition was compared to a previously used composition. A reaction time of 30 minutes was found optimal for the new CuAAC composition, improving the old composition regarding reaction rate. Both compositions showed great labeling yields in HBSS with no labeling in complete DMEM. When reducing concentrations of toxic agents in the new CuAAC reaction, labeling yields only slightly decreased when one of the toxic agent concentrations was halved. The composition with halved CuSO4 concentration showed low specificity in live labeling which excluded the composition from further research. A MTT assay showed the highest cell viabilities when treating cells with the new CuAAC reaction composition with halved NaAsc concentrations.
Item Type:Essay (Bachelor)
Faculty:TNW: Science and Technology
Programme:Biomedical Technology BSc (56226)
Link to this item:https://purl.utwente.nl/essays/92269
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