By D.B. Roodyn (Eds.)
Within the face of fast advancements in automatic innovations, this ebook provides a very good consultant to offer tendencies. It presents info on: ideas and terminology of enzyme automation; automated tools illustrated by way of the Technicon approach; semi-automatic equipment; interrupted-flow and discrete-sampling platforms; single-enzyme research; multiple-enzyme research (M.E.A.); enzyme characterization; calculation of enzyme actions from device readings; generalized platforms for enzyme automation; Appendices I. released automatic enzyme assays, II. Terminology utilized in enzyme automation, III. equipment utilized in enzyme automation, IV. laptop software for generalized enzyme computerized approach.
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If a spectrophotometer with an automatic cell changer is used with 4 cuvettes) but in general the method can only operate a t a rate of analysis comparable to continuous-flow methods if the time of incubation is small. Where the actual rate of analysis is not a critical factor, clearly measurement of the entire progress curve on a stationary reaction mixture has great advantages. Continuous monitoring of enzyme activities is difficult to perform with interrupted-flow systems because of the intermittent flow of enzyme.
One must also consider the possibility of surface denaturation of enzyme during the assay. It is also difficult to automate methods in which the enzyme preparation is very viscous, for example with bacterial lysates containing much DNA. It is therefore necessary to consider each case on its own merits, but in the light of the general principles indicated above. 1. Interrupted-flow systems We have seen in ch. 2 how the semi-automatic methods rely on manual loading of the autosensor. , the reaction mixture is prepared, mixed with enzyme and pumped into the sensor, usually the flow cell of a recording spectrophotometer.
If it takes one minute to pipette the sample and reaction mixture into a test-tube, about 100 min may be spent preparing racks of tubes manually. These could all be incubated simultaneously (perhaps for 5 min). If the tubes could be read manually at a rate of one every 30 Eec, the entire manual assay could take about half the timc of the semi-automatic assay. If the rate of reaction is slow, and it takes 10-20 min to obtain a satisfactory reading, the difference in time becomes even greater. To overcome this difficulty, manufacturers of recording spectrophotometers have introduced automatic sample changers of various degrees of complexity.
Automated Enzyme Assays by D.B. Roodyn (Eds.)