{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:32:22Z","timestamp":1760239942876,"version":"build-2065373602"},"reference-count":30,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2019,1,28]],"date-time":"2019-01-28T00:00:00Z","timestamp":1548633600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>Back scatter interferometry (BSI) is a sensitive method for detecting changes in the bulk refractive index of a solution in a microfluidic system. Here we demonstrate that BSI can be used to directly detect enzymatic reactions and, for the first time, derive kinetic parameters. While many methods in biomedical assays rely on detectable biproducts to produce a signal, direct detection is possible if the substrate or the product exert distinct differences in their specific refractive index so that the total refractive index changes during the enzymatic reaction. In this study, both the conversion of glucose to glucose-6-phosphate, catalyzed by hexokinase, and the conversion of adenosine-triphosphate to adenosine di-phosphate and mono-phosphate, catalyzed by apyrase, were monitored by BSI. When adding hexokinase to glucose solutions containing adenosine-triphosphate, the conversion can be directly followed by BSI, which shows the increasing refractive index and a final plateau corresponding to the particular concentration. From the initial reaction velocities, KM was found to be 0.33 mM using Michaelis\u2013Menten kinetics. The experiments with apyrase indicate that the refractive index also depends on the presence of various ions that must be taken into account when using this technique. This study clearly demonstrates that measuring changes in the refractive index can be used for the direct determination of substrate concentrations and enzyme kinetics.<\/jats:p>","DOI":"10.3390\/s19030539","type":"journal-article","created":{"date-parts":[[2019,1,29]],"date-time":"2019-01-29T03:40:55Z","timestamp":1548733255000},"page":"539","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Real-Time Interferometric Refractive Index Change Measurement for the Direct Detection of Enzymatic Reactions and the Determination of Enzyme Kinetics"],"prefix":"10.3390","volume":"19","author":[{"given":"S\u00f8ren T.","family":"Jepsen","sequence":"first","affiliation":[{"name":"Department of Clinical Biochemistry, Aalborg University Hospital, Sdr. Skovvej, DK-9000 Aalborg, Denmark"},{"name":"Department of Clinical Medicine, Aalborg University, Sdr. Skovvej, DK-9000 Aalborg, Denmark"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2920-1343","authenticated-orcid":false,"given":"Thomas M.","family":"J\u00f8rgensen","sequence":"additional","affiliation":[{"name":"Department of Applied Mathematics and Computer Science, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark"}]},{"given":"Henrik S.","family":"S\u00f8rensen","sequence":"additional","affiliation":[{"name":"Department of Photonics Engineering, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark"}]},{"given":"S\u00f8ren R.","family":"Kristensen","sequence":"additional","affiliation":[{"name":"Department of Clinical Biochemistry, Aalborg University Hospital, Sdr. Skovvej, DK-9000 Aalborg, Denmark"},{"name":"Department of Clinical Medicine, Aalborg University, Sdr. Skovvej, DK-9000 Aalborg, Denmark"}]}],"member":"1968","published-online":{"date-parts":[[2019,1,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"4855","DOI":"10.3390\/s100504855","article-title":"A comprehensive review of glucose biosensors based on nanostructured metal-oxides","volume":"10","author":"Rahman","year":"2010","journal-title":"Sensors"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1038\/nrd838","article-title":"Optical biosensors in drug discovery","volume":"1","author":"Cooper","year":"2002","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"664","DOI":"10.1007\/s00216-002-1235-9","article-title":"Fiber-optic biosensors\u2014An overview","volume":"372","author":"Marazuela","year":"2002","journal-title":"Anal. Bioanal. 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