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ELISA stages

Shkedy, Z. et al., Modeling anti-KLH ELISA data using two-stage and mixed effects models in support of immunotoxicological studies, J. Biopharm. Stat., 15, 205, 2005. [Pg.17]

Biosensors Biosensors are used in the final measurement stage of immunoassays such as an electrochemistry-based enzyme-linked immunoassay (ELISA) or the measurement of catalyse-labelled antigens at an antibody-coated oxygen electrode. [Pg.93]

At an early stage in product development, there must be some material made available for use as ELISA reagents. ELISA requires a standard for quantitation and specific antibodies to the drug protein of interest. If these are not already available, either commercially or internally, then time, effort, and expense must be reserved for reagent development as well as assay development activities. The specific antibody is the key reagent in an ELISA. The antibody defines the specificity and sensitivity of the assay. To date there is no successful substitution for the routine production of specific antibodies by immunization of an animal with the antigen target of interest. [Pg.281]

ELISAs can be used for identification and quantitation of a biopharmaceutical product or for quantitation of impurities or contaminants as discussed previously. They can be used throughout the manufacturing process as well as in quality control or the product release stage just as they are used in all the other stages of product development. To be used for quality control, GMP practices must be followed. All methods need to be validated so that the assay s performance is documented. ELISAs should have internal quality controls to monitor assay... [Pg.294]

Both the ELISA and Western blot suffer from the problem that antibodies may not appear in an aposed individual s blood until months after the initial exposure. Methods for using PCR to screen blood samples for HIV are being developed, PCR amplification of the HIV provirai DNA 7-ovides the ability to detect HIV at earlier stages of infection, because the viral nucleic acid is r resent immediately upon exposure. It is used to detect HIV infection in newborns whose mothers are HIV positive. [Pg.107]

ELISA has been used for measuring caspase activity. For the ELISA of intracellular caspase activity at the very early stages of apoptosis, apoptotic cells are first lysed to isolate their intracellular contents. Different caspase activities in the cell lysate can then be determined by the addition of a caspase-specific tetrapeptide substrate that is conjugated to the color reporter molecule p-nitroanilide (pNA) (e.g., DEVD-pNA for caspase-3 and lETD-pNA for caspase-8). The cleavage of the substrate peptide by the caspase releases the chromophore pNA, which can be... [Pg.90]

Fecal elastase 1 concentration can be measured by an ELISA test kit using an antibody specific for the human enzyme pancreatin supplements do not interfere with this pancreatic function test and need not be discontinued. Although measurement of fecal elastase 1 excretion appears to be somewhat more sensitive than fecal chymotrypsin, its specificity and positive predictive value are similarly low, and falsepositive results can be expected in patients with intestinal diseases. Conversely, mild-to-moderate stages of pancreatic exocrine insufficiency cannot be diagnosed reliably. [Pg.284]

The initial emphasis in analytical biotechnology was on broad safety concerns that translated into detection of host-cell components such as DNA, endotoxins, Escherichia colt proteins, and retroviral contamination.2 The detection of these impurities requires development of high-sensitivity assays that are based primarily on antibodies [e.g., enzyme-linked immunosorbent assay (ELISA) for E. coli proteins) or radioactivity (e.g., dot-blot assays for DNA detection). New developments are focused on low-sensitivity detection, characterization, and removal of undesirable target sequence variants. Bioseparations play an important role even after a product has been isolated and shown to contain a low level of contaminants for initiation of clinical studies. The focus shifts to achievement of a reproducible, large-scale manufacturing process. At this stage, analytical methods provide essential informa-... [Pg.694]

FIGURE 45.2. IL-6 secretion from myoblasts, fusing myoblasts, and myotubes. Cultured human myoblasts, fusing myoblasts, and myotubes were added DFP at the 10 M concentration. The secreted IL-6, calculated per 100,000 nuclei, was determined 24 h later with ELISA kit (Endogen, Rockford, IL, USA). Significant difference (Student I-test, p < 0.05 n — 3) between control and treated cultures was observed at all stages studied but was most prominent in the myoblasts. [Pg.685]


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See also in sourсe #XX -- [ Pg.45 , Pg.82 ]




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