Enzyme immunoassay techniques

An enzyme immunoassay technique has been employed for measuring endosulfan and its degradation products (i.e., endosulfan diol, endosulfan sulfate, endosulfan ether, and endosulfan lactone) in water at 3 ppb (Chau and Terry 1972 Musial et al. 1976). However, this technique is not currently in use in environmental residue analysis. Further research into this technique could produce a rapid, rehable, and sensitive method for identifying contaminated areas posing a risk to human health. No additional methods for detecting endosulfan in environmental media appear to be necessary at this time. However, methods for the determination of endosulfan degradation products are needed. 

All specimens were analyzed by EMIT (Enzyme Immunoassay Technique), an enzyme method based upon the competitive bonding of an enzyme and an antibody. This method yields a positive result with concentrations of 75 ng of PCP/ml or greater (Rubenstein et al. 

Polynuclear aromatic hydrocarbons (PAH) are aromatic compounds that contain two or more benzene rings fused together. These substances may be analyzed by HPLC, GC, GC/MS and enzyme immunoassay techniques. The latter is a rapid screening method that may be applied for a qualitative or semiquanti-tative determination Test kits are commercially available for such screening. U.S. EPA (1995) has specified a method (Draft Method 4035) that detects a range of PAHs to different degrees and measures the composite of individual responses to determine the total PAHs in the sample. 

L9. Litman, D. J., Hanlon, T. M., and Ullman, E. F., Enzyme channeling immunoassay A new homogeneous enzyme immunoassay technique. Anal. Biochem. 106, 223-229... 

The enzyme immunoassay technique described below is a simple and sensidve method to detect left-handed Z-DNA formation in synthedc poly-nucleoddes, recombinant plasmids, and nadve DNAs. This method udlizes the differences in the andgenic properdes of right-handed B-DNA and left-handed Z-DNA. 

The enzyme immunoassay technique consists of five sequential steps ... 

This relationship is established by measurement of samples with known amounts of analyte (calibrators). One may distinguish between solutions of pure chemical standards and samples with known amounts of analyte present in the typical matrix that is to be measured (e.g., human serum). The first situation applies typically to a reference measurement procedure, which is not influenced by matrix effects, and the second case corresponds typically to a field method that often is influenced by matrix components and so preferably is calibrated using the relevant matrix. Calibration functions may be linear or curved, and in the case of immunoassays often of a special form (e.g., modeled by the four-parameter logistic curve) This model (logistic in log x) has been used for both radioimmunoassay and enzyme immunoassay techniques and can be written in several forms as shown (Table 14-1). Nonlinear regression analysis is applied to estimate the relationship, or a logit transforma-... 

T Porstmann, ST Kiessig. Enzyme immunoassay techniques, an overview. J Immunol Meth 150 5, 1992. 

BC Sallustio, et al. Unbound plasma phenytoin concentrations measured using enzyme immunoassay technique on the Cobas Mira Analyzer—In vivo effect of valproic acid. Ther Drug Monit 14 9, 1992. 

J.P. Sherry, A. Borgmann, Enzyme-immunoassay Techniques for the Detection of Atrazine in Water Samples - Evaluation of a Commercial Tube Based Assay , Ghemosphere, 26, 2173-2184 (1993). 

Interference with diagnostic tests A 96-year-old woman with delirium had a false-positive urine screen for opiates after starting levofloxacin [39" ]. An enzyme immunoassay technique (SYNCHRON) was used initially, but repeat testing of the urine and serum by mass spectroscopy was negative. Enzyme... 

Because of this lack of resolving power, much electroanalytical research is aimed at providing increased selectivity. This can be accomplished in two ways. First, electrochemistry can be combined with another technique which provides the selectivity. Examples of this approach are liquid chromatography with electrochemical detection (LCEC) and electrochemical enzyme immunoassay (EEIA). The other approach is to modify the electrochemical reaction at the electrode to enhance selectivity. This... 

Several heterogeneous electrochemical enzyme immunoassays have been demonstrated. These are based on the enzyme-linked immunosorbent assay (ELISA) technique... 

Additionally it has been our experience that mass spectrometry as a routine detection/identification technique for bacteria is not well received by microbiologists and clinicians who prefer less expensive, less complicated approaches to bacterial typing and identification, such as methods based on polymerase chain reaction (PCR) and enzyme-linked immunosorbent assays (ELISA). For that reason we have adapted our MS approach to serve as a means of biomarker discovery that feeds candidate proteins or leads into development as PCR targets or other immunoassay techniques. 

As an alternative, extremely sensitive detection can be achieved with reporter antibody probes tagged with intensely SERS-active compounds or with enzymes that react with substrates to yield SERS-active products. These methods often involve sandwich immunoassay techniques, which increase the number of required steps but offer the advantages of excellent sensitivity and the potential for label multiplexing. For example, Nie and coworkers recently reported the simultaneous detection of two types of antigens in a... 

Alternative methods of diagnosis include enzyme immunoassay, DNA probes, and nucleic acid amplification techniques. 

Pankey et al.21 described a rapid, reliable, and specific enzyme multiplied immunoassay technique (EMIT ) for amitriptyline, nortriptyline, imipramine, and desipramine in sera. To overcome crossreactivity, solid phase extraction was included in sample pretreatment. Disposable 1 mL columns packed with covalently labeled silica gel were conditioned with HPLC-grade methanol (1 mL) and then with de-ionized or distilled water (1 mL). Serum (calibrator, control, or patient sample, 500 L) was applied onto the column, eluted to waste, washed with 900 /uL of wash solution containing acetonitrile (236.1 g/L) and ion-pairing reagent in acetate buffer, pH 4.2, washed with 500 fiL of mobile phase solution containing acetonitrile (393.5 g/L) in methanolic phosphate buffer, pH 7.0,... 

With enzyme-multiplied immunoassay technique (EMIT) assays, enzyme tags are used instead of radiolabels. The antibody binding alters the enzyme characteristics,... 

Electrophoretic and isotachoelectrophoretic techniques are gaining in popularity in soil analysis with applications to polyaromatic hydrocarbons, polychlorobiphenyls, tetrahydrothiophene and triazine herbicides, Paraquat and Diquat and growth regulators. Other lesser-used techniques include spectrophotometric methods (five determinants), spectrofluorimetric methods (two determinants), luminescence methods (one determinant), titration methods (one determinant), thin-layer chromatography (five applications), NHR spectroscopy (two applications) and enzymic immunoassays (one determinant). 

The introduction of enzyme immunoassay (EIA) and similar allied immunoassay techniques in early eighties showed, in fact, a brighter path towards quantitative analysis. 

The most common of these systems is the enzyme-multiplied immunoassay technique or EMIT, which is particularly suited to the measurement of small molecules (haptens) such as drugs. EMIT is a trade mark of the Syva Corporation of Palo Alto, California. Although it does not involve the separation of bound fraction from free it is nevertheless a competitive assay system. The antigen is labelled with an enzyme in such a way that the enzyme retains its catalytic activity. When the antigen binds to the antibody the enzyme becomes inhibited, probably by an induced conformational change or by steric hindrance of the enzyme active site (Figure 7.15). 

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