Monoclonal enzyme

Abad, A., M.J. Moreno, R. Pelegrf, et al. 1999. Determination of carbaryl, carbofuran and methiocarb in cucumbers and strawberries by monoclonal enzyme immunoassays and high-performance liquid chromatography with fluorescence detection An analytical comparison. J. Chromatogr. A 833 3-12. 

The first assay (a radioimmunoassay) that measured cTnl used polyclonal anti-cTnl antibodies. The first monoclonal enzyme-linked immunosorbent assay, anti-cTnl antibody-based immunoassay, was described by Bodor and co-workers (1992). Numerous manufacturers have now developed monoclonal antibody-based diagnostic immunoassays for the measurement of cTnl in serum. Assay times range from 5 to 30 minutes. 

Chen C, Malone KE, Prunty J, Daling JR. Measurement of urinary estrogen metabolites using a monoclonal enzyme-linked immunoassay kit Assay performance and feasibility for epidemiological studies. Cancer Epidemiol Biomarkers Prev 1996 5 727-32. 

Menendez-Botet, C. J., Stankievic, R., Schwartz, D., and Schwartz, M. K., Comparison of the quantitative measurement of human estrogen and progesterone receptor in tissue cytosol by the dextran-coated charcoal assay and the Abbott monoclonal enzyme immunoassay. Clin. Chem. (Winston-Salem, N.C.) 34(6), 1225 (1988). 

Wl. Weigand, R. A., Cotter, D. L., Dunn, R. A., Nolan, C., Greene, G., and Przywara, L. W., Quantitation of progesterone receptor (PgR) in human breast tumors by double monoclonal enzyme immunoassay. Breast Cancer Res. Treat. 8(1), 87 (1976). 

R.J.A. Deschamps, J.C. Hall, M.R. McDermott, Polyclonal and Monoclonal Enzyme Immunoassays for Picloram Detection in Water, Soil, Plants and Urine , J. Agric. Food Chem., 38,1881 (1990). 

A two-site immunometric assay of undecapeptide substance P (SP) has been developed. This assay is based on the use of two different antibodies specifically directed against the N- and C-terminal parts of the peptide (95). Affinity-purified polyclonal antibodies raised against the six amino-terminal residues of the molecule were used as capture antibodies. A monoclonal antibody directed against the carboxy terminal part of substance P (SP), covalently coupled to the enzyme acetylcholinesterase, was used as the tracer antibody. The assay is very sensitive, having a detection limit close to 3 pg/mL. The assay is fiiUy specific for SP because cross-reactivity coefficients between 0.01% were observed with other tachykinins, SP derivatives, and SP fragments. The assay can be used to measure the SP content of rat brain extracts. 

Mammalian Cells Unlike microbial cells, mammalian cells do not continue to reproduce forever. Cancerous cells have lost this natural timing that leads to death after a few dozen generations and continue to multiply indefinitely. Hybridoma cells from the fusion of two mammalian lymphoid cells, one cancerous and the other normal, are important for mammalian cell culture. They produce monoclonal antibodies for research, for affinity methods for biological separations, and for analyses used in the diagnosis and treatment of some diseases. However, the frequency of fusion is low. If the unfused cells are not killed, the myelomas 1 overgrow the hybrid cells. The myelomas can be isolated when there is a defect in their production of enzymes involved in nucleotide synthesis. Mammahan cells can produce the necessary enzymes and thus so can the fused cells. When the cells are placed in a medium in which the enzymes are necessaiy for survival, the myelomas will not survive. The unfused normal cells will die because of their limited life span. Thus, after a period of time, the hybridomas will be the only cells left ahve. 

Bioprocess plants are an essential part of food, fine chemical and pharmaceutical industries. Use of microorganisms to transform biological materials for production of fermented foods, cheese and chemicals has its antiquity. Bioprocesses have been developed for an enoimous range of commercial products, as listed in Table 1.1. Most of the products originate from relatively cheap raw materials. Production of industrial alcohols and organic solvents is mostly originated from cheap feed stocks. The more expensive and special bioprocesses are in the production of antibiotics, monoclonal antibodies and vaccines. Industrial enzymes and living cells such as baker s yeast and brewer s yeast are also commercial products obtained from bioprocess plants. 

Enzyme inhibitors Monoclonal antibodies Steroids Vaccines... 

Enzyme-Mediated Substrate Immunolocalization ofPoIygalacturonic Acid Within Barley Epidermal Cell Walls Utilizing Endopolygalacturonase of Cochliobolus sativus and a Monoclonal Antibody Specific for the Enzyme... 

Figure 1. Transverse section of barley leaf epidermal cells taken perpendicular to the long axis of the cells and anticlinal to the leaf surface. The section has been labeled by the EMSIL technique (see Methods) utilizing purified C. sativus endopolygalacturonase and monoclonal antibody EPG-4, which is specific for this enzyme, in order to localize the substrate of the enzyme at the typical site penetrated by the fungal pathogen. Bar = 1 pm. Inset Comparable cell wall region as in Fig. 1 but labeled with monoclonal antibody JIM 5 to localize non-esterified pectin. Bar = 1 pm. Note the identical labeling patterns obtained with either method.

The approximate location of the epitopes for more than 40 monoclonal anti-ATPase antibodies has been mapped to various regions within the cytoplasmic domain of the Ca " -ATPase [285,302-304]. All antibodies were found to bind with high affinity to denatured Ca -ATPase, but the binding to the native enzyme showed significant differences depending on the location of antigenic sites within the ATPase molecule. 

Bonfante et al. (73) used monoclonal antibodies and enzyme-gold complexes to reveal pectins and cellulose at the interface between the fungal wall and the host plasma membrane in AM roots (Fig. 6), and additional wall components have been investigated with other molecular probes (74-76). These studies indicate that the interface is an apoplastic space of high molecular complexity where the boundaries of the partners are defined. The examination of other endomycorrhizal systems has demonstrated that their interface is morphologically similar but different in composition. Cellulose and pectins are present at the interface... 

Watanabe et al. developed an enzyme-linked immunosorbent assay (ELISA) for the detection of inabenfide, a plant growth regulator, in rice. Specific monoclonal antibody (MAB) is used for this method. The effects of rice matrices on the sensitivity of ELISA can be reduced by adding 0.1% Tween 20. Good reproducibility and accuracy of the proposed ELISA were obtained for rice samples and the recovery was 92% at a fortification level of 5-500 xgkg . ... 

Muldoon et al. developed a monoclonal-based competitive inhibition enzyme-linked immunosorbent assay (cELISA) for sulfadimethoxine. The group compared... 

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