ELISA analysis

Enzyme - linked immnnosorbent analysis (ELISA) is a method known for decades. The technology is based on the ability of a specific antibody to distinguish the tri-dimensional structure of a given mycotoxin. The direct competitive... 

Sample preparation techniques vary depending on the analyte and the matrix. An advantage of immunoassays is that less sample preparation is often needed prior to analysis. Because the ELISA is conducted in an aqueous system, aqueous samples such as groundwater may be analyzed directly in the immunoassay or following dilution in a buffer solution. For soil, plant material or complex water samples (e.g., sewage effluent), the analyte must be extracted from the matrix. The extraction method must meet performance criteria such as recovery, reproducibility and ruggedness, and ultimately the analyte must be in a solution that is aqueous or in a water-miscible solvent. For chemical analytes such as pesticides, a simple extraction with methanol may be suitable. At the other extreme, multiple extractions, column cleanup and finally solvent exchange may be necessary to extract the analyte into a solution that is free of matrix interference. 

Immunoassays designed for environmental applications are mostly sold as some variation of the ELISA format. ELISA-like formats dominate the field because they are inexpensive and because they provide high sensitivity and precision without requiring complex instrumentation. The basic ELISA format supports both field and laboratory-based applications but is limited by multiple steps and inadequate sensitivity for some applications, excessive variability and sometimes long analysis times. Some of the other formats discussed in this article may replace the ELISA for selected applications however, because many laboratories are familiar with the ELISA technology, there will be a significant delay before alternative formats are widely accepted. 

Oxime carbamates are not directly amenable to gas chromatography (GC) because of their high thermal instability, which often leads to their breakdown at the injection port or in the column during analysis. Analysis of oxime carbamates by GC with sulfur detection or flame photometric detection involves oxidation of the intact insecticides or alkaline hydrolysis to form the more volatile but stable oxime compound. Enzymatic techniques have been reported for the analysis of these compounds. Enzyme-linked immunosorbent assay (ELISA) has been used to determine aldicarb and its sulfone and sulfoxide metabolites and methomyl in water, soil, and sediment samples. 

In the last decade, modem biochemical methods have been used for analysis of protein binders [20,21] in one case a group led by A. Heginbotham identified egg proteins in a seventeenth century painting using immunofluorescent microscopy and enzyme-linked immunosorbent assay (ELISA) [20]. 

Starodub et al. [98] studied different constructions and biomedical applications of immunosensors based on fiberoptic and enhanced CL. They discussed three different approaches of immobilization of one of the immunocomponents on the fiberoptic surface. Good results could be achieved by the use of a special membrane closely connected to the fiberoptic, with sensitivities compared to that obtained by the ELISA method but with a faster rate of analysis. The sensor was... 

Abstract A significant number of immunochemical methods have been described for the determination of the most important emerging pollutants. The present chapter is a compilation of the information available today regarding immunochemical determination of industrial residues with a high potential risk of causing negative effects in the environment, wildlife, and public health. Homogeneous immunoassays, ELISAs, FIIAs, immunosensors, and selective immunoaffinity sample treatment methods have been reported for the analysis of an important number of these substances. The bases of these methods are briefly presented. 

Several qualitative and quantitative immunochemical methods for CAP analysis in biological matrices of animal origin have been described [101,102, 104,105] (see Table 3). Van de Water et al. [ 102] described an ELISA that detected CAP in swine muscle tissue with an IC50 value of 3 ng mL1. This immunoassay was improved and subsequently optimized incorporating the streptavidin-biotin amplification system. There are also several commercially available test kits (see Table 4). RIDASCREEN is a competitive enzyme immunoassay for the quantitative analysis of CAP residues in milk, eggs, and meat in a microtiter plate. The measurement is made photometrically, obtaining a LOD of 100 ng L 1 in meat and eggs and 150 ng L 1 in milk. The test has been also applied to the analysis of tetracyclines. 

Immunochemical methods have been developed and placed on the market to analyze tetracycline residues (see Table 4). Thus, a qualitative EIA has been developed and used to analyze tetracyclines in honey samples with a detection level of 20 pg/kg-1 [96]. A microplate-based indirect ELISA has been developed to analyze tetracyclines using polyclonal antibodies. The assay could measure tetracycline in the range between 0.1 and 6 ng mL L Other tetracycline antibiotics such as chlortetracycline, rolitetracycline, or minocycline are also highly recognized in this assay [98]. Several immunoassay kits are commercially available for the analysis of tetracyclines although, to our knowledge, none of them... 

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