Assay Biochemistries

Melki, R., Fievez, S., and Carlier, M.-F. (1996). Continuous monitoring of Pi release following nucleotide hydrolysis in actin or tubulin assembly using 2-amino-6-mer-capto-7-methylpurine ribonucleoside and purine-nucleoside phosphorylase as an enzyme-linked assay. Biochemistry 35, 12038-12045. 

As mentioned above, the central core component of a genotyping technology is contained within the assay biochemistry. In this regard, a general distinction may be made between those methods that rely primarily upon differential hybridization stringency for their specificity, and those that derive specificity primarily from the ability to detect a product of an enzymatic reaction. A number of... 

Schwartz B, Markwalder JA, Seitz SP, Wang Y, Stein RL. A kinetic characterization of the glycosyltransferase activity of Eschericia coli PBP lb and development of a continuous fluorescence assay. Biochemistry 2002 41 12552-12561. 

Ozers, M.S., Marks, B.D., Gowda, K., Kupcho, K.R., Ervin, K.M., De Rosier, T., Qadir, N., Eliason, H.C., Riddle, S.M. and Shekhani, M.S. (2007) The androgen receptor T877A mutant recruits LXXLL and FXXLF peptides differently than wild-type androgen receptor in a time-resolved fluorescence resonance energy transfer assay. Biochemistry, 46, 683-695. 

Godemann R, Madden J, Kramer J et al (2009) Fragment-based discovery of BACE1 inhibitors using functional assays. Biochemistry 48 10743-10751... 

Hey, T., Lipps, G., Sugasawa, K., Iwai, S., Hanaoka, F., and Krauss, G. (2002). The XPC-HR23B complex displays high affinity and specificity for damaged DNA in a true equilibrium fluorescence assay. Biochemistry 41, 6583-6587. 

Mofron. J.L. Kuzmic. P. Kishore, V. Colonbonilla. E. Rich. D.H. Determination of kinetic constants for peptidyl prolyl cis trails isomerases by an improved spectrophoto-metric assay. Biochemistry 1991. 30, 6127-6134. 

Stegmann, T., Schoen, P., Bron, R., Wey, J., Bartoldus, I., et al. Evaluation of viral membrane fusion assays. Comparison of the octadecylrhodamine dequenching assay with the pyrene excimer assay. Biochemistry 32, 11330-11337 (1993)... 

Aeed, P. A. Sperry, A. E. Young, C. L. Nagiec, M. M. EUiammer, A. P. Effect of membrane perturbants on the activity and phase distribution of inositol phosphorylceramide synthase development of a novel assay. Biochemistry 2004, 43, 8483-8493. 

Stults, N. L., et al. (1992). Use of recombinant biotinylated aequorin in microtiter and membrane-based assays Purification of recombinant aequorin from Escherichia coli. Biochemistry 31 1433-1442. 

The following brief account identifies only major groups of herbicides not mentioned elsewhere in the text, and is far from comprehensive. Their mode of action is only dealt with in a superficial way. From an ecotoxicological point of view, there has not been as much concern about their sublethal effects upon plants as there has been in the case of mammals, and there has not been a strong interest in the development of biomarker assays to establish their effects. The major concern has been whether weeds, or nontarget plants, have been removed following herbicide application—a rather easy matter to establish as plants are fairly sedentary. For a more detailed account of herbicide chemistry and biochemistry, see Hassall (1990). 

The author gratefully acknowledges the cooperation of Dr. Roger Cooke, the Department of Biochemistry and Biophysics, U.C.S.F., for contractile protein-related materials and assays. 

So far, we have reviewed the various ways in which complex dose-response curves in intact-tissue bioassays can be the result, the pharmacological resultant, of two or more interacting activities. Now, if all that these bioassays achieved was to blur and obscure the underlying activities, they would have to give way to the newer, analytically simpler assays based on chemistry and biochemistry. However, the beauty of intact-tissue bioassays is that they are analytically tractable by using families of dose-response curves and appropriate mathematical models, the complexity of intact hormone-receptor systems can, indeed, be interpreted. Bioassay allows them to be studied as systems in ways denied to simple biochemical assays. 

In carrying out an enzyme assay it may be convenient to introduce an auxiliary enzyme to the system to effect the removal of a product produced by the first enzymatic reaction. McClure [Biochemistry, 8 (2782), 1969] has described the kinetics of certain of these coupled enzyme assays. The simplest coupled enzyme assay system may be represented as... 

Gorun, V., Proinov, I., Baltescu, V., Balaban, G. and Barzu, O. (1978). Modified Ellman procedure for assay of cholinesterases in crude enzymatic preparations. Analytical Biochemistry 86 324-326. 

Silvius, J.R., Leventis, R., Brown, P.M., and Zuchermann, M. (1987) Novel fluorescent phospholipids for assays of lipid mixing between membranes. Biochemistry 26, 4279-4287. 

Sturgeon, C. and McAllister, E. 1998. Analysis of hCG clinical applications and assay requirements. Annals of Clinical Biochemistry 35, 460-491. 

J.E. Fairbrother in "Assay of Drugs and Other Trace Compounds in Biological Fluids, Ed. by E. Reid, "Methodological Developments in Biochemistry, Vol. 5, Longman Group, London, 1976, p. 141. 

Ward, A. M. Catto, J. W. F. Hamdy, F. C., Prostate specific antigen Biology, biochemistry and available commercial assays, Ann. Clin. Biochem. 2001, 38, 633 651... 

Dashek WV, Micales J. Assay and purification of enzymes-oxalate decarboxylase, in Methods in Plant Biochemistry and Molecular Biology (Dashek WV, ed.), CRC Press, Boca Raton, FL, 1997, pp. 49-71. 

The occurrence of more than one affected child in many of the families with galactosemia made it appear highly probable that this was a genetically determined disease. The mode of inheritance was worked out by the conventional methods of human genetics and, later, confirmed by biochemical studies of the families. The direct assay of enzymatic activity in heretozygotes for galactosemia must rank as one of the major contributions of biochemistry to the study of human genetics. 

Biology and biochemistry laboratories perform three general types of assays ... 

Crespi, C.L., Miller, V.P. and Penman, B.W. (1997) Microtiter plate assays for inhibition of human, drug-metabolizing cytochromes P450. Analytical Biochemistry, 248 (1), 188-190. 

Biomedical analytical chemistry happens to be one of the latest disciplines which essentially embraces the principles and techniques of both analytical chemistry and biochemistry. It has often been known as clinical chemistry . This particular aspect of analytical chemistry has gained significant cognizance in the recent past by virtue of certain important techniques being included very much within its scope of analysis, namely colorimetric assays, enzymic assays, radioimmunoassays and automated methods of clinical analysis. 

Enzymes occupy an important place in analytical biochemistry and many investigations require their detection and quantitation. Studies of the enzyme content of blood plasma are particularly useful in clinical biochemistry both in the monitoring of normal metabolic processes and in the detection of abnormal levels of enzyme production or release. Enzyme assays also provide convenient methods for assessing the quality of foodstuffs and checking the efficiency of sterilization and pasteurization processes. 

A family of 100 hybridoma antibodies can typically provide 20 tight binders and these need to be assayed for catalysis. At this stage in the production of an abzyme, the benefit of a sensitive, direct screen for product formation comes into its own. Following identification of a successful catalyst, the antibody is usually recloned to ensure purity and stabilization of the clone, then protein is produced in larger amount (—10 mg) and used for determination of the kinetics and mechanism of the catalysed process by classical biochemistry. Digestion of such protein with trypsin or papain provides fragment antibodies, Fabs, that contain only the attenuated upper limbs of the intact IgG (Fig. 1). It is these components that have been crystallized, in some... 

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