Automated kinase assays

Creatine kinase (CK) -clinical assay for [AUTOMATED INSTRUMENTATION - CLINICALCHEMISTRY] (Vol3)... 

EEC assays can be easily automated and miniaturized due to their homogeneous format and can therefore be used to screen large libraries of chemical compounds against a large diversity of biological targets, such as kinases or G Protein Coupled Receptors. 

The high FRET efficiency obtained in HTRF assays enables a large variety of biological events to be probed. Protein/protein interactions, enzymatic activities (e.g. kinases or proteases), or a large number of biomarkers have been detected with a very low detection limit. Like the other homogeneous fluorescent technologies like FP or FCS, HTRF can be easily automated and miniaturized down to a 1536 weUs plate format for the HTS of large libraries of chemical compounds. ... 

ST53 Butch, A.W., Goodnow, T.T., Brown, W.S., McCellan, A., Kessler, G. and Scott, M.G. (1989). Stratus automated creatine kinase-MB assay evaluated Identification and elimination of falsely increased results associated with a high-molecular-mass form of alkaline phosphatase. Clin. Chem. 35, 2048-2053. 

The development of automated luminometers is focused primarily on devices that achieve high sample throughput rates, typically through the use of 96 and 384 well microtitre plates. Whilst suited for many applications, these systems require a skilled operator and often do not provide rapid results when the time taken to prepare the microtitre plate is included in the assay time. An instrument that can perform on demand, automated, near real time analysis using a variety of luminescent assay protocols has been developed. The instrument has been designed to automate various luminescent assays including adenylate kinase (AK) assays and those that use magnetic separation steps in conjunction with bioluminescence. ... 

Previously, we tested a manual method using antibiotic-mediated lysis of nontarget cells followed by immuno-magnetic separation with an adenylate kinase (AK) bioluminescence endpoint determination. This 4 h assay for patient swabs was carried out on a limited number of samples. The work reported here was to introduce automation into the assay and to carry out tests on a larger number of samples in a hospital setting. Initial assay development was done in a non-clinical laboratory using spiked samples. The assay was then adapted to fit in with the standard hospital test. Modifications were introduced as testing in the hospital laboratory proceeded. 

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