文章基本信息

标题：Monitoring extracellular ion and metabolite dynamics with recombinant nanobody-fused biosensors
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作者：Sandra Burgstaller ; Teresa R. Wagner ; Helmut Bischof 等
期刊名称：iScience
印刷版ISSN：2589-0042
出版年度：2022
卷号：25
期号：9
页码：1-22
DOI：10.1016/j.isci.2022.104907
语种：English
出版社：Elsevier
摘要：SummaryIon and analyte changes in the tumor microenvironment (TME) alter the metabolic activity of cancer cells, promote tumor cell growth, and impair anti-tumor immunity. Consequently, accurate determination and visualization of extracellular changes of analytes in real time is desired. In this study, we genetically combined FRET-based biosensors with nanobodies (Nbs) to specifically visualize and monitor extracellular changes in K+, pH, and glucose on cell surfaces. We demonstrated that these Nb-fused biosensors quantitatively visualized K+alterations on cancer and non-cancer cell lines and primary neurons. By implementing a HER2-specific Nb, we generated functional K+and pH sensors, which specifically stained HER2-positive breast cancer cells. Based on the successful development of several Nb-fused biosensor combinations, we anticipate that this approach can be readily extended to other biosensors and will open new opportunities for the study of extracellular analytes in advanced experimental settings.Graphical abstractDisplay OmittedHighlights•Generation of recombinant nanobody-fused FRET biosensors•Nb-fused biosensors specifically bind targets on the outer surface of various cells•Cellular bound Nb-biosensors visualize extracellular analyte changes in real timeSensor; Optical imaging; Biochemistry