Advertisement

Separation-Free Enzyme Immunoassay for Haptens

  • Anna Jaklitsch

Abstract

Separation-free (homogeneous) enzyme immunoassays (Emit®) were first developed at the Syva Research Institute in the early 1970s. Their discovery was that binding of an anti-drug antibody to an enzyme labeled with the same drug, modulated (inhibited or activated) catalytic activity. Thus a competitive immunoassay can be developed in which drug in the sample and enzyme-labeled drug compete for antibody. Modulation of enzyme activity is directly related to the analyte concentration in the sample and is monitored without separation of antibody bound and unbound fractions. This principle was later extended to a variety of enzymes and also applied to the measurement of hormones (Ullman et al., 1979) and proteins (Gibbons et al., 1980).

Keywords

Therapeutic Drug Monitoring Dextran Sulfate Enzyme Conjugate Toxicology Assay Oxidize nicotinamIde Adenine Dinucleotide 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bach, P.R. and J.W. Larsen (1980). Stability of standard curves prepared for EMIT homogeneous enzyme immunoassay kits stored at room temperature after reconstitution, Clin. Chem. 26: 652–654.PubMedGoogle Scholar
  2. Carrico, R.J., J.E. Christner, R.C. Boguslaski and K.K. Yeung (1976). A method for monitoring specific binding reactions with cofactor labeled ligands, Anal. Biochem. 72; 271–282.PubMedCrossRefGoogle Scholar
  3. Centofanti, J., A. Monte, D. Grandsaert, B. Chung, M. Ruggieri, R. Schneider, and D. Kabakoff (1981). The EMIT®-stTM drug detection system, J. Clin. Chem. Clin. Biochem. 19: 631.Google Scholar
  4. Chang, J., S. Gotcher, J.B. Gushaw (1982). Homogeneous enzyme immunoassay for theophylline in serum and plasma, Clin. Chem. 28: 361–367.PubMedGoogle Scholar
  5. Cox, C. and J. Buret (1982). Spectrophotometry homogeneous immunoassay of serum gentamicin with oxidized nicotinamide adenine dinucleotide as label, Anal. Chem. 54: 1862–1865.PubMedCrossRefGoogle Scholar
  6. Deleide, V., V. Dona, and R. Malvano (1979). Homogeneous enzyme-immunoassay for anticonvulsant drugs: Effects of hapten-enzyme bridge length, Clin. Chem. Acta 99: 195–201.CrossRefGoogle Scholar
  7. Dols, J.L. and A.P. van Zanten (1981). Gentamicin assay: comparison of an adapted EMIT method and an RIA method, Ann. Clin. Biochem. 18: 236–239.PubMedGoogle Scholar
  8. Fukuchi, H., M. Yoshida, S. Tsukiai, T. Kitaura, and T. Konishi (1984). Comparison of enzyme immunoassay, radioimmunoassay, and microbiologic assay for amikacin in plasma, Am. J. Hosp. Pharm. 41: 690–693.PubMedGoogle Scholar
  9. Gibbons, I., C. Skold, G.L. Rowley, and E.F. Ullman (1980). Homogeneous enzyme immunoassay for proteins employing β-galacto-sidase, Anal. Biochem. 102: 167.PubMedCrossRefGoogle Scholar
  10. Gorsky, J.E., P.R. Bach, and W. Wong (1983). Improved sensitivity of aminoglycoside enzyme immunoasays, Clin. Chem. 29: 1994–1995.PubMedGoogle Scholar
  11. Gorsky, J.E. (1983). EMIT assay curve stability, Clin. Chem. 29: 1690.PubMedGoogle Scholar
  12. Hamlin, C.R., and P.A. Sullivan (1984). Adaptation of EMIT reagents to the Cobas Bio centrifugal analyzer, Clin. Chem. 30: 314–315.PubMedGoogle Scholar
  13. Iosefsohn, M., R.L. Boeckx, and J.M. Hicks (1981). Adaptation of the EMIT gentamicin and tobramycin procedure to the IL micro-centrifugal analyzer, Ther. Drug Mon. 3: 365–370.CrossRefGoogle Scholar
  14. Jaklitsch, A.P., R.S. Schneider, R.J. Johannes, J.E. Lavine, and G.L. Rosenberg (1976). Homogeneous enzyme immunoassay for T4 in serum, Clin. Chem. 22: 1185.Google Scholar
  15. Jaklitsch, A.P., C.B. Gagne, D.S. Kabakoff, D.K. Leung, P. Singh, and W. Protzmann (1980). Measurement of sisomicin concentration in fermentation broths by homogeneous enzyme immmunoassay, Am. Chem. Soc. Meeting MBT 23, Las Vegas, Nev., American Chemical Society.Google Scholar
  16. Krogsted, D.J., G.G. Granich, P.R. Murray, M.A. Pfaller, and R. Valdes (1982). Heparin interferes with the radioenzymatic and homogeneous enzyme immunoassays for aminoglycosides, Clin. Chem. 28: 1517–1521.Google Scholar
  17. Lockwood, W.R. and J.D. Bower (1973). Tobramycin and gentamicin concentrations in the serum of anephric patients, Antimicrob. Agents Chemother. 3: 125–129.PubMedGoogle Scholar
  18. Matzke, G.R., C. Gwizdala, J. Wery, D. Ferry, and R. Starnes (1982). Evaluation of three gentamicin serum assay techniques, Ther. Drug Mon. 4: 195–200.CrossRefGoogle Scholar
  19. Matzke, G.R., R. Piveral, C.E. Halstenson, and P.A. Abraham (1984). Heparin interferes with the tobramycin serum concentration determinations by EMIT, Drug Intell. Clin. Pharm. 18: 517–519.PubMedGoogle Scholar
  20. Ngo, T.T. and H.M. Lenhoff (1980). Enzyme modulators as tools for the development of homogeneous enzyme immunoassays, FEBS Lett. 116: 285–288.PubMedCrossRefGoogle Scholar
  21. Oellerich, M., R. Haeckel, and H. Haindl (1982). Evaluation of EMIT adapted to the (Cobas) Bio centrifugal analyzer, J. Clin. Chem. Clin. Biochem. 20: 765–772.PubMedGoogle Scholar
  22. Ou, C.N., V.L. Frawley, and G.J. Buffone (1981). Optimization of EMIT reagent system using a Cobas-Bio centrifugal analyzer, J. Anal. Toxicol. 5: 249–252.PubMedGoogle Scholar
  23. Ratcliff, R.M., C. Mirelli, E. Moran, D. O’Leary, and R. White (1981). Comparison of five methods for the assay of serum gentamicin, Antimicrob. Agents Chemother. 19: 508–512.PubMedGoogle Scholar
  24. Rowley, G.L., K.E. Rubenstein, S.T. Weber, and E.F. Ullman (1976). Mechanism by which antibodies inhibit hapten-glucose-6-phosphate dehydrogenase, Am. Chem. Soc, Meeting BIOL 151, Washington, D.C., American Chemical Society.Google Scholar
  25. Rubenstein, K.E., R.S. Schneider, and E.F. Ullman (1972). “Homogeneous” enzyme immunoassay. A new immunochemical technique, Biochem. Biophys. Res. Commun. 47: 846–851.PubMedCrossRefGoogle Scholar
  26. Singh, P.S., D.K. Leung, G.L. Rowley, C. Gagne, and E.F. Ullman (1980). A method for controlled coupling of amino compounds to enzymes: a homogeneous enzyme immunoassay for gentamicin, Anal. Biochem. 104: 51–58.PubMedCrossRefGoogle Scholar
  27. Tindula, R.J., P.J. Ambrose, and A.F. Harralson (1983). Aminoglycoside inactivation by penicillins and cephalosporins and its impact on drug-level monitoring, Drug Intell. Clin. Pharm. 17: 906–908.PubMedGoogle Scholar
  28. Ullman, E.F., R.A. Yoshida, J.I. Blakemore, W. Eimsted, R. Ernst, R. Leute, D.K. Leung, E.T. Maggio, R. Ronald, and B. Sheldon (1979). Mechanism of inhibition of malate dehydrogenase by thyroxine derivatives and reactivation by antibodies. Basis for a homogeneous enzyme immunoassay for thyroxine, Biochim. Biophys. Acta. 567: 66–74.PubMedGoogle Scholar
  29. Wellington, D. (1981). Mathematical treatments for the analysis of enzyme-immunoassay data. In Enzyme-Immunoassay (E.T. Maggio, ed.) CRC Press, Boca Raton, Fla.Google Scholar
  30. Whiting, B., A.W. Kelman, S.M. Bryson, F.H.M. Derkx, A.H. Thomson, G.H. Fotheringham, and S.E. Joel (1984). Clinical Pharmacokinetics: a comprehensive system for therapeutic drug monitoring and prescribing, Br. Med. J. 288: 541–545.CrossRefGoogle Scholar
  31. Witebsky, F.G., C.A. Suva, S.T. Selepak, M.E. Ruddel, J.D. MacLowry, E.E. Johnson, and R.J. Elin (1983). Evaluation of four gentamicin and tobramycin assay procedures for clinical laboratories, J. Clin. Microbiol. 18: 890–894.PubMedGoogle Scholar
  32. Woo, J., M.A. Longley, and D.C. Cannon (1982). Homogeneous enzyme immunoassay for tobramycin evaluated and compared with a radioimmunoassay, Clin. Chem. 28: 1370–1374.PubMedGoogle Scholar
  33. Wu, J.W., C. Bunyagidj, S. Hoskin, S.M. Riebe, J. Aucker, K. White, and S.P. O’Neill (1983). Quantitation of haptens by homogeneous immunoprecipitation. 2. Centrifugal analysis for tobramycin, phénobarbital, and theophylline in serum, Clin. Chem. 29: 1540–1542.PubMedGoogle Scholar
  34. Wu, J.W., S. Hoskin, S.M. Riebe, J.E. Gifford, and S.P. O’Neill (1982). Quantitation of haptens by homogeneous immunoprecipitation. 1. Automated analysis of gentamicin in serum, Clin. Chem. 28: 659–661.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • Anna Jaklitsch
    • 1
  1. 1.SYVA CompanyPalo AltoUSA

Personalised recommendations