Immunochemical analysis method

In the third part, methods for retrospective detection of exposure to toxic scheduled chemicals using mass spectrometric and immunochemical analysis methods are discussed. The described methods are applied to human origin samples. These methods are essential when in cases of use, or allegations of use, previous presence or absence of toxic chemicals need to be confirmed. Identification of CWC-related chemicals provides key supporting evidence of noncompliance with the CWC. 

Oudin, J., Specific precipitation in gels and its application to immunochemical analysis. Methods in Med. Research 5, 335 (1952). 

Oudin, J. (1980) Immunochemical analysis by antigen-antibody precipitation in gels. Methods Enzymol. 70,166-198. 

Protein quantification is an important step for handling protein samples for isolation and characterization, and is a prerequisite step before submitting proteins for chromatographic, electrophoretic, or immunochemical analysis and separation. The methods included in this unit are colorimetric measurements, whose procedures are faster, simpler, and less laborious than those based on estimation of total nitrogen content (vnitbi.2). 

Nichkova, M. and M.R Marco. 2005. Development and evaluation of C18 and immunosorbent solid-phase extraction methods prior to immunochemical analysis of chlorophenols in human urine. Anal. Chim. Acta 533 67-82. 

Standardization of immunochemical methods will be needed for their widespread usage in different countries and laboratories. This will include SOPs (standard operating procedures), GLP (good laboratory practice), and participation in proficiency tests (PTs) that will be comparable to those carried out routinely for conventional analysis, but with adaptations to tailor them for immunochemical analysis. 

Noort D, Black RM. Methods for the retrospective detection of exposure to toxic scheduled chemicals. Part B mass spectrometric and immunochemical analysis of covalent adducts to proteins and DNA. In Mesilaakso M, ed. Chemical Weapons Convention Chemicals Analysis Sample Collection. Preparation, and Analytical Methods, Chichester, West Sussex, England John Wiley Sons 2005 433 51. 

Immunochemical methods are rapidly gaining acceptance as analytical techniques for pesticide residue analysis. Unlike most quantitative methods for measuring pesticides, they are simple, rapid, precise, cost effective, and adaptable to laboratory or field situations. The technique centers around the development of an antibody for the pesticide or environmental contaminant of interest. The work hinges on the synthesis of a hapten which contains the functional groups necessary for recognition by the antibody. Once this aspect is complete, immunochemical detection methods may take many forms. The enzyme-linked immunosorbent assay (ELISA) is one form that has been found useful in residue applications. This technique will be illustrated by examples from this laboratory, particularly molinate, a thiocarbamate herbicide used in rice culture. Immunoassay development will be traced from hapten synthesis to validation and field testing of the final assay. 

Matrix effects in food samples frequently occur owing to colored extracts or to the content of hpids, proteins or polyphenols that may be coextracted during sample preparation. As food samples usually have to be extracted prior to immunochemical analysis, the method of analyte extraction is of great importance. Analytes... 

Subsequent to the RIA development an ELISA method for EO exposure monitoring, was develop. The ELISA method was more rapid and convenient than the RIA, retaining the advantages of immunochemical analysis but without the hazards of radioactivity. 

Only limited development of new methodologies has taken place for immunochemical analysis of nucleic acids. Most published methods rely on modifications to classical DNA probe hybridization or immunoassay methods, with considerable blending of the two. For example, some methods employ immobilized oligonucleotide probes to capture the analyte DNA followed by immunoenzymatic detection. Other methods use immunocapture followed by detection with an enzyme-labeled DNA probe. Distinctly new methodologies mostly impact on assay formats (e.g., DNA microarrays and in situ hybridization) and detection reagents (e.g., chemiluminescent enzyme substrates). 

Another major asset in the routine purification and analysis of the eicosanoids is high-performance liquid chromatography (HPLC). In particular, reversed-phase HPLC has proven to be extremely effective, especially when combined with radioactive, spectroscopic or immunochemical detection methods.For example, the combination of reversed-phase HPLC with ultraviolet absorbance spectroscopy was instrumental to the discovery of leukotrienes and lipoxins, and it is still widely used to detect these compounds as well as other ultraviolet-absorbing eicosanoids. ... 

J. M. Van Emon and R. O. Mumma, Immunochemical Methods for Environmental Analysis, ACS Symposium Series 442, American Chemical Society, Washington, D.C., 1990. 

N. Haagsma and C. van der Water, Immunochemical methods in the analysis of veterinary drug residues, in Analysis of Antibiotic Drug Residues in Food Products of Animal Origin, ed. V. K. Agarwal, Plenum Press, New York, pp. 81-97 (1992). 

PC. Peng, S.J. Wratten, E.W Logusch, S.R. Horton, and R.C. Sharp, in ACS Symposium Series, Immunochemical Methods in Environmental Analysis, American Chemical Society, Washington, DC, Chapter 15 (1990). 

Van Emon, J. M. Immunochemical Methods of Environmental Analysis. Washington, DC American Chemical Society, 1989. 

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