By D.B. Roodyn (Eds.)
Within the face of speedy advancements in automatic innovations, this publication provides a superb consultant to offer tendencies. It presents details on: ideas and terminology of enzyme automation; automated tools illustrated by way of the Technicon process; semi-automatic tools; interrupted-flow and discrete-sampling platforms; single-enzyme research; multiple-enzyme research (M.E.A.); enzyme characterization; calculation of enzyme actions from software readings; generalized structures for enzyme automation; Appendices I. released computerized enzyme assays, II. Terminology utilized in enzyme automation, III. gear utilized in enzyme automation, IV. machine application for generalized enzyme automatic method.
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Additional resources for Automated Enzyme Assays
However, in a general sense it is any tube, probe or pump that connects a given reagent to the analytical system. For automation, we therefore use the terms SubjecI index p . 218 42 AUTOMATED ENZYME ASSAYS line dilution factor and fractional line volume. For convenience these may be abbreviated to LDF and FLV respectively. 3. Adaptation to the autoanalyzer Let us now see how the above simple assay may be adapted to the autoanalyzer. A simple flow-system to perform the assay is shown in fig. 1. Phosphate and lactate are pumped into the system and mixed in a mixing coil (MC,).
Another important method of studying enzyme-catalyzed reactions is by measuring changes in fluorescence. In his excellent book on ‘Fluorescence Assay in Biology and Medicine’, Udenfriend (1962) discusses the use of such measurements in enzymology and demonstrates their widespread application and great sensitivity. Fluorimetry is also used to measure common substrates and co-factors, by the use of specific enzymes. As with light absorption methods, there is a great range in sophistication of the instruments used.
A larger number of samples can be assayed at the same time. Temperature control is rigorous and it is possible to agitate and gas the vessels during the course of the reaction. It is interesting how fashion may sometimes dictate the equipment purchased. Many ingenious and fruitful techniques of manometry were developed before 1939 to follow enzyme reactions, for example using bicarbonate buffers in a COz gas phase to monitor acid-base changes (Dixon 1951). These methods are now generally neglected in favour of the recording spectrophotometer.