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S. Armenta Estrela, W. Tomischko, B. Lendl:
"A mid-IR flow-through sensor for label free monitoring of enzyme inhibition";
Applied Spectroscopy, 62 (2008), 12; 1322 - 1325.

Label free monitoring of acetylcholinesterase (AChE) activity was achieved with a mid-IR flow through sensor. The flow-through sensor comprised agarose beads, carrying covalently immobilized AChE, which were placed in a temperature controlled (37°C) CaF2 flow cell with an optical path of 60 µm. The sensor was incorporated into a computer controlled sequential injection (SI) system for automated liquid handling. Different mixtures of enzyme substrate acetylcholine (ACh) and inhibitor (tacrine) were prepared and fed into the flow-through sensor. The flow was stopped as soon as the prepared mixtures had reached the sensor. Enzymatic hydrolysis of ACh by AChE was directly monitored as it took place in the flow-through sensor. The inhibition effect of tacrine was calculated from the reaction induced spectral changes revealing an important decrease in the activity of AChE, approaching zero when the inhibitor concentration is high enough. The developed mid-IR flow-through sensor is flexible and can be used to study the inhibitor activity of different target molecules as well as different enzymes

Flow-through sensor, FTIR, SIA, enzyme, inhibition kinetics, acetylcholinesterase

http://publik.tuwien.ac.at/files/PubDat_167179.pdf