Enzyme immunoassay method development

Schuetz [191] has developed an enzyme immunoassay method for the determination of all polychlorobiphenyls up to biphenyl in soils before determination by ELISA. 

Brandon [11] developed an enzyme immunoassay method using monoclonal antibodies to determine thiabendazole in apples and potatoes. 

Harrison, R. O. Brimfield, A. A. Nelson, J. O. Development of a monoclonal antibody based enzyme immunoassay method for analysis of maleic hydrazide. J. Agric. Food Chem. 37 958-964. 1989. 

Liu F, Liu FH, Zhuo RX et al. Development of a polymer-enzyme immunoassay method and its application. Biotechnol Appl Biochem 1995 21 257-264. 

Assays of ciguatoxin. Determination of ciguatoxin levels in fish was carried out in many laboratories by mouse assays. Enzyme immunoassay to screen inedible fish has been proposed by Hokama (9). No specific chemical assay has been developed, as information on functional groups suitable for fluorescence labeling is not available. Analyses conducted in the authors laboratory on remnant fish retrieved from patients meals indicated that ciguatoxin content as low level as 1 ppb could cause intoxication in adults. An extremely high sensitivity and a sophisticated pretreatment method will be required for designing a fluorometric determination method for the toxin. 

Using a simple solvent extraction procedure to minimize matrix effects, a diclofop-methyl immunoassay was developed for milk, a number of edible plant products, and other matrices. Gas chromatography (GC) and liquid scintillation counting (LSC) of a C-labeled analyte were used as reference methods to compare with enzyme immunoassay (EIA) results. The methods were well correlated, with comparison of EIA... 

L/mole). It cross-reacts at >90% with saxitoxin but at <1% with neosaxitoxin. This antibody, when used in an anti-rabbit IgG "second antibody" radioimmunoassay format, can detect pmole quantities of saxitoxin. This assay has been shown to be a simple and efficient method for the analysis of saxitoxin in clam extracts. The lack of antibody cross-reactivity to the neosaxitoxin sub-group of the paralytic shellfish poisons limits the general utility of the assay to neurophysiology studies and to certain clam species which preferentially accumulate saxitoxin. However, the radioimmunoassay serves as a good precursor in the development of an enzyme immunoassay for the paralytic shellfish poisons. 

Barbour, H. M. (1976) Development of an enzyme immunoassay for human placental lactogen using labelled antibodies./. Immunol. Methods 11, 15. 

As mentioned earlier, the immunoassay can be done with a number of procedural modifications and in this instance one must substitute the isotope with another molecule (fluorescent dye, magnetic particle, enzyme) which can be measured and therefore serve as the source of tracer. For our initial studies we have chosen to use the enzyme immunoassay (EIA) system. At the present time the EIA is still in its infancy and although a number of successful EIA s have been developed the method cannot be considered a panacea (34). The future of this assay appears to be very bright and exciting, and there is considerable interest in the application of the EIA to problems in both microbiology and clinical medicine (34). Many of the procedures and protocols are derived from RIA procedures and the EIA, like the RIA, has the potential to be performed in a multitude of procedural variations but, for the purpose of this manuscript we will describe only the system we have chosen for our use. 

Immunoassay methods use antibodies developed against one or more triazines and enzymes that create a colored signal. The result is a sensitive, simple method of detecting triazines in a wide variety of matrices. An immunoassay requires significant resources to develop, but once developed can provide rapid, inexpensive analyses that can be specific to a single triazine or sensitive to a variety of triazines. 

Chemiluminescence immunoassay methods have many applications (Weeks, 1992). Highly sensitive chemiluminescent immunoassays were developed by Tsuji et al. (1989) for determination of enzymes (oxidases, peroxidase, glucose oxidase, P-D-galactosidase) as well as various hormones and drugs in biological fluids (Tsuji et al., 1989). 

Aizawa et al. reported the development of a competitive enzyme immunoassay using electrodes (A2). In this method, antibody immobilized on the membrane is attached to the electrodes for oxygen. When catalase-antigen conjugate binds to the immobilized antibody, production of oxygen can be... 

In 1976, we developed a double-antibody enzyme immunoassay of TSH using alkaline phosphatase, but this method was not very sensitive (M8). As shown in Table 10, several other methods were later reported (A3, 12, K2, K3, M7) and two methods have been used to measure TSH in dried blood samples in screening for neonatal hypothyroidism in Japan. 

Atopic diseases including atopic dermatitis and bronchial asthma develop during the infantile period and are found to be predictable at birth by detecting a high immunoglobulin E (IgE) concentration in cord blood. Croner et al. reported a radioimmunoassay of IgE for screening for these disorders (C8). A method of enzyme immunoassay of IgE in serum has been developed (H6) and may be used for this purpose. 

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