Enzyme-linked immunosorbent assay monoclonal antibodies

Watanabe et al. developed an enzyme-linked immunosorbent assay (ELISA) for the detection of inabenfide, a plant growth regulator, in rice. Specific monoclonal antibody (MAB) is used for this method. The effects of rice matrices on the sensitivity of ELISA can be reduced by adding 0.1% Tween 20. Good reproducibility and accuracy of the proposed ELISA were obtained for rice samples and the recovery was 92% at a fortification level of 5-500 xgkg . ... 

Valdivieso-Garcia, A. Riche, E. Abubakar, O. Waddell, T. E. Brooks, B. W. A double antibody sandwich enzyme-linked immunosorbent assay for the detection of Salmonella using biotinylated monoclonal antibodies. J. Food Prot. 2001, 64,1166-1171. 

King, D.P. et al., The use of monoclonal antibodies specific for seal immunoglobulins in an enzyme-linked immunosorbent assay to detect canine distemper virus-specific immunoglobulin in seal plasma samples, J. Immuno. Methods, 160, 163, 1993. 

Enzyme labels are usually associated with solid-phase antibodies in the technique known as enzyme-linked immunosorbent assay (ELISA). There are several variants of this technique employing both competitive and non-competitive systems. However it is best used in combination with two monoclonal antibodies in the two-site format in which an excess of antibody is bound to a solid phase such as a test-tube or microtitre plate the test antigen is then added and is largely sequestered by the antibody (Figure 7.12). After washing... 

LI. Labeur, G., Michiels, G., Bury, J., Usher, D. C., and Rosseneu, M., Lipoprotein(a) quantified by an enzyme-linked immunosorbent assay with monoclonal antibodies. Clin. Chem. 

W18. Wong, W. L. T., Eaton, D. L., Berloui, A., Fendly, B., and Hass, P. E., A monoclonal-antibody-based enzyme-linked immunosorbent assay for lipoprotein(a). Clin. Chem. (Winston-Salem. NC) 36, 192-197 (1990). 

D. J. Bucher, A. Mikhail, S. Popple, P. Graves, G. Meiklejohn, D. S. Hodes, K. Johansson, andP. E. Halonen, Rapid detection of Type A Influenza viruses with monoclonal antibodies to the M protein (Ml) by enzyme-linked immunosorbent assay and time-resolved fluoroimmunoassay, J. Clin. Microbiol. 29, 2484-2488 (1991). 

Enzyme-linked immunosorbent assay (ELISA) Immnne-based quantitation of expressed protein Rapid, specifle, quantitative, and efficient Requires one or more high-afflnity antisera or monoclonal antibodies... 

The assessment of DNA adducts may provide a sensitive indicatCH of previous exposure. The enzyme-linked immunosorbent assay (ELISA) has a lower limit of detection of about 0.08 femtomol per microgram of DNA (Perera et oL, 1982). This assay requires (1) the development of an antibody specific for a certain chemical metabolite bound covalently to DNA and (2) the isolation of DNA from some tissue sample, ag., skin biopsy, or lymphocytes of an exposed individual. It is anticipated that further refinement of such immunologic techniques may lower the threshold of sensitivity by one or two orders of magnitude. One such refined test is the ifitrasensitive ymatic radioimmunoassay (USE-RIA), purported to be about five times more sensitive than ELISA (Hsu et oL, 1981 Shamsuddin et oL, 1985 Harris et oL, 1985). Quantification by the development of monoclonal antibodies to aflatoxin Bj metabolites bound to DNA (Groopman et oL, 1982 Sizaret et oL, 1982) has now been reported. 

Kobori K, Saito K, Ito S, Kotani K, Manabe M, Kanno T (2002) A new enzyme-linked immunosorbent assay with two monoclonal antibodies to specific epitopes measures human lecithin-cholesterol acyltransferase. J Lipid Res 43 325-334... 

Immunoaffinity cleanup was first applied in drug residue analysis for the determination of chloramphenicol in swine muscle tissue by LC (113). The lAC column was prepared using monoclonal antibodies originally developed for an enzyme-linked immunosorbent assay (ELISA) method (171) specific for chloramphenicol. Meat samples were extracted with water, and a concentrated phosphate buffer was added to the filtered extracts before immunoaffinity cleanup. A phosphate buffer was used in the washing process, whereas chloramphenicol was eluted from the column with a glycine/sodium chloride solution of pH 2.8. For subsequent LC analysis, this eluate was extracted with ethyl acetate, evaporated, and reconstimted in the mobile phase. The same analytical scheme was later successfully applied for the determination of chloramphenicol in eggs and milk as well (170, 172). 

Enzyme-linked immunosorbent assay (ELISA) is a very useful technique for the specific and sensitive assay of certain compounds, in which suitable antibodies, monoclonal or polyclonal, to the compounds are available. The technique has found particular application m the monitoring of environmental contaminants and toxins, either studying the primarily contaminated materials, e.g., foodstuffs, or body fluids of potentially exposed humans. The technique has been increasingly applied to monitoring the carcinogenic mycotoxins, the aflatoxins. 

I Barna-Vetro, A Gyoengyoesi, L Solti. Monoclonal antibody-based enzyme-linked immunosorbent assay of fusarium T-2 and zearalenone toxins in cereals. Appl Env Microbiol 60(2) 729-731, 1994. 

Antibody Assays Monoclonal antibodies can be analyzed by the solid-phase assay techniques such as enzyme linked immunosorbent assays (ELISA) or radioimmuno assays (RIA). The typical assay procedures are as follows (Figure 5.9)... 

Test wells exhibiting growth for the presence of monoclonal antibodies when the hybridoma growth covers approx 25% of the base of the cell well. The assay system used should mimic the final test format required. Commonly, Triple Antibody Sandwich (TAS) (5) enzyme-linked immunosorbent assay (ELISA) is use for screening hybridomas for specific antibody. It is important when testing hybri-... 

Lucas, A.D., P. Schneider, R.O. Harrison, J.N. Seiber, B.D. Hammock, J.W. Biggar, and D.E. Rolston (1991). Determination of atrazine and simazine in water and soil using polyclonal and monoclonal antibodies in enzyme-linked immunosorbent assays. Food Agric. Immunol., 3 155-167. 

Goda, Y., M. Hirobe, A. Kobayashi, et al. 2005. Production of a monoclonal antibody and development of enzyme-linked immunosorbent assay for alkyl ethoxylates. Anal. Chim. Acta 528 47-54. 

Competitive binding immunoassay (e.g., RIA) Enzyme-linked immunosorbent assay (ELISA) Immunoradiometric assay (IRMA) — dual monoclonal antibody assay Receptor binding assay Cell binding assay... 

An enzyme-linked immunosorbent assay (ELISA) was developed by Van der Schans and colleagues for detection of the mustard adduct within DNA, using monoclonal antibodies raised against N7-HETE-guanosine-5 -phosphate coupled to keyhole limpet hemocyanin (n). The ELISA was successfully applied in toxicokinetic studies in which levels of adducted DNA were followed in conjunction with measurement of intact sulfur mustard (12). 

The first assay (a radioimmunoassay) that measured cTnl used polyclonal anti-cTnl antibodies. The first monoclonal enzyme-linked immunosorbent assay, anti-cTnl antibody-based immunoassay, was described by Bodor and co-workers (1992). Numerous manufacturers have now developed monoclonal antibody-based diagnostic immunoassays for the measurement of cTnl in serum. Assay times range from 5 to 30 minutes. 

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