Enzyme Immunoassay of Plant Constituents

  • Richard L. Mansell
  • Cecilia A. McIntosh


The use of immunoassay to quantitate plant derived compounds was first developed in clinical laboratories in the early 1970’s. These first assays were directed toward small organic molecules, e. g., colchicine (Boudene et al., 1975), nicotine (Langone et al., 1973), and morphine (Spector, 1971), which have medicinal or pharmaceutical value. These assays were based on the radioimmunoassay (RIA) and were developed for compounds whose small molecular size renders them non-immunogenic. Assays for low molecular weight compounds of animal origin had been developed earlier (see Erlanger, 1980, and references therein) using synthesized protein-hapten conjugates for immunization. The immunoassays mentioned above demonstrated that the hapten-protein conjugate principle (Landsteiner, 1945) also could be used to induce animals to produce antibodies against compounds of plant origin.


Abscisic Acid Soya Protein Enzyme Immunoassay Antibody Preparation Lysergic Acid 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Aldwell, F.E.B., Hall, I.R. and J.M.B. Smith. 1985. Enzyme-linked immunosorbent assay as an aid to taxonomy of the Endogonaceae. Trans. Br. Mycol. Soc. 84: 399–402.CrossRefGoogle Scholar
  2. Arens, H. and M.H. Zenk. 1980. Radioimmunoassays for the determination of lysergic acid and simple lysergic acid derivatives. Planta Medica 39: 336–347.CrossRefGoogle Scholar
  3. Atzorn, K. and E.W. Weiler. 1983. The immunoassay of gibberellins. II. Quantitation of GA3, GA4, and GA7 by ultra-sensitive enzyme immunoassays. Planta 159: 7–11.CrossRefGoogle Scholar
  4. Barthe, G.A. and I. Stewart. 1985. Enzyme immunoassay (EIA) of endogenous cytokinin in Citrus. J. Agric. Food Chem. 33: 293–297.CrossRefGoogle Scholar
  5. Bolwell, G.P., Bell, J.N., Cramer, C.L., Schuch, W., Lamb, C.J. and R.A. Dixon. 1985. L-phenylalanine ammonia lyase from Phaseolus vulgaris — characterization and differential induction of multiple forms from elicitor-treated cell suspensions. Eur. J. Biochem. 149: 411–419.PubMedCrossRefGoogle Scholar
  6. Borrebeack, C.A.K. 1984. Detection and characterization of a lectin from non-seed tissues of Phaseolus vulgaris. Planta 161: 223–228.CrossRefGoogle Scholar
  7. Borrebaeck, C.A.K. and B. Mattiasson. 1983. Distribution of a lectin in tissues of Phaseolus vulgaris. Physiol. Plant. 58: 29–32.CrossRefGoogle Scholar
  8. Borrebeack, C.A.K. and M.E. Etzler. 1981. Production and characterization of a monoclonal antibody against the seed lectin of the Dolichos biflorus plant. J. Biol. Chem. 256: 4723–4725.Google Scholar
  9. Boudene, C., Duprey, F. and C. Bohuon. 1975. Radioimmunoassay of colchicine. Biochem. J. 151: 413–415.PubMedGoogle Scholar
  10. Campbell, W.H. and K.G. Ripp. 1984. An elisa for higher plant nitrate reductase. Annals N.Y. Acad. Sci. 435: 123–125.CrossRefGoogle Scholar
  11. Conroy, J.M. and A. Esen. 1984. An enzyme-linked immunosorbent assay for zein and other proteins using unconventional solvents for antigen adsorption. Anal. Biochem. 137: 182–187.PubMedCrossRefGoogle Scholar
  12. Daie, J. and R. Wyse. 1982. Adaptation of the enzyme-linked immunosorbent assay (ELISA) to the quantitative analysis of abscisic acid. Anal. Biochem. 119: 365–371.PubMedCrossRefGoogle Scholar
  13. Eberle, J., Arnscheidt, A., Klix, D. and E.W. Weiler. 1986. Monoclonal antibodies to plant growth regulators III. zeatinriboside and dihydrozeatinriboside. Plant Physiol. In Press.Google Scholar
  14. Engvall, E. and P. Perlmann. 1971. Enzyme-linked immunosorbent assay (ELISA) quantitative assay of immunoglobulin G. Immunochem. 8: 871–874.CrossRefGoogle Scholar
  15. Engvall, E. and A.J. Pesce. 1978. Quantitative enzyme immunoassay. Suppl. #7, Scan. J. Immunol. Blackwell Scientific Publications, (Oxford). 129 pp.Google Scholar
  16. Erlanger, B.F. 1980. The preparation of antigenic hapten-carrier conjugates: a survey. in Methods in Enzymology, ed. by H. van Vunakis and J.J. Langone. Academic Press, N.Y. vol 70(a): 85–104.Google Scholar
  17. Gold, P., Lewis, M., Mazzarella, R. and M. Green. 1985. An enzyme-linked immunoassay for the detection of antibodies to endoplasmic reticulum. Anal. Biochem. 146: 82–89.PubMedCrossRefGoogle Scholar
  18. Griffiths, N.M., Billington, M.J., Crimes, A.A. and C.H.S. Hitchcock. 1984. An assessment of commercially available reagents for an enzyme-linked immunosorbent assay of soya protein in meat products. J. Sci. Food Agric. 35: 1255–1260.CrossRefGoogle Scholar
  19. Hansen, C.E., Wenzler, H. and F. Meins, Jr. 1984. Concentration gradients of trans-zeatin riboside and trans-zeatin in the maize stem. Plant Physiol. 75: 959–963PubMedCrossRefGoogle Scholar
  20. Higgins, R.C. and L.D. Gottlieb. 1984. Subunit hybridization and immunological studies of duplicated phosphoglucose isomerase isozymes. Biochem. Genet. 22: 957–979.PubMedCrossRefGoogle Scholar
  21. Hirai, M. 1983. Seasonal changes in sorbitol-6-phosphate dehydrogenase in loquat leaf. Plant & Cell Physiol. 24: 925–931.Google Scholar
  22. Hitchcock, C.H.S., Bailey, F.J., Crimes, A.A., Dean, D.A.G. and P.J. Davis. 1981. Determination of soya proteins in food using an enzyme-linked immunoassay procedure. J. Sci. Food Agric. 32: 157–165.CrossRefGoogle Scholar
  23. Horgen, P.A., Nakagawa, C.H. and R.T. Irvin. 1984. Production of monoclonal antibodies to a steroid plant growth regulator. Can. J. Biochem. Cell Biol. 62: 715–721.CrossRefGoogle Scholar
  24. Hunt, R.E. and L.H. Pratt. 1979. Phytochrome radioimmunoassay. Plant Physiol. 64: 327–331.PubMedCrossRefGoogle Scholar
  25. Jordan, B.R., Partis, M.D. and B. Thomas. 1984. A study of phytochromemembrane association using an enzyme-linked immunosorbent assay and western blotting. Physiol. Plant. 60: 416–421.CrossRefGoogle Scholar
  26. Jourdan, P.S., E.W. Weiler and R.L. Mansell. 1985. Naringin levels in citrus tissues. I. Comparison of different antibodies and tracers for the radioimmunoassay of naringin. Plant. Physiol. 77: 896–902.PubMedCrossRefGoogle Scholar
  27. Jourdan, P.S., Mansell, R.L., Oliver, D.G. and E.W. Weiler. 1984. Competitive solid phase enzyme-linked immunoassay for the quantification of limonin in Citrus. Anal. Biochem. 138: 19–24.PubMedCrossRefGoogle Scholar
  28. Jourdan, P.S., Weiler, E.W. and R.L. Mansell. 1983. Radioimmunoassay for naringin and related flavanone 7-neohesperidosides using a tritiated tracer. J. Agric. Food Chem. 31: 1249–1255.CrossRefGoogle Scholar
  29. Jourdan, P.S., Mansell, R.L. and E.W. Weiler. 1982. Radioimmunoassay for the citrus bitter principle, naringin, and related flavonoid-7-0-neo-hesperidosides. Planta Med. 44: 82–86PubMedCrossRefGoogle Scholar
  30. Kanoaka, M., Yano, S., Kato, H., Nakanishi, K. and M. Yoshizaki. 1984. Studies on the enzyme immunoassay of bio-active constituents contained in Oriental medicinal drugs. III. Enzyme immunoassay of paeoniflorin, a constituent of Chinese paeony root. Chem. Pharm. Bull. 32: 1461–1466.CrossRefGoogle Scholar
  31. Kanoaka, M., Yano, S., Kato, H., Nakao, N. and E. Kinoshita. 1983. Studies on the enzyme immunoassay of bio-active constituents contained in Oriental medicinal drugs. II. Enzyme immunoassay of glycyrrhizin. Chem. Pharm. Bull. 31: 1866–1873.CrossRefGoogle Scholar
  32. Kanoaka, M., Yano, S., Kato, H. and N. Nakano. 1981. Glycyrrhetylamino acids: synthesis and application to enzyme immunoassay for glycyrrhetic acid. Chem. Pharm. Bull. 29: 1533–1538.CrossRefGoogle Scholar
  33. Kosfizer, M., Clausell. A., Imam, S., Lavery Jr., B., Border, B., Witte, P. and W.J. Snell. 1982. A monoclonal antibody that blocks adhesion of mt+ gametes of Chlamydomonas. J. Cell Biol. 95: 71a.Google Scholar
  34. Landsteiner, K. 1945. The specificity of serological reactions. Harvard University Press, (Cambridge, MA).Google Scholar
  35. Langone, J.J., Gjika, H.B. and H. Van Vunakis. 1973. Nicotine and its metabolites. Radioimmunoassays for nicotine and cotinine. Biochem. 12: 5025–5030.CrossRefGoogle Scholar
  36. Maggio;, E.T. 1980. Enzyme-immunoassay. CRC Press, Inc.(Boca Raton, FL). 295 pp.Google Scholar
  37. Maier, V.P., Hasegawa, S., Bennett, R.D., and L.C. Echols. 1980. Limonin and limonoids: chemistry, biochemistry, and juice bitterness. in Citrus nutrition and quality, ed. by S. Nagy and J.A. Attaway. ACS Symposium Series 143: 63–82.CrossRefGoogle Scholar
  38. Maier, V.P., Bennett, R.D. and S. Hasegawa. 1977. Limonin and other limonoids. in Citrus science and technology, ed. by S. Nagy, P.E. Shaw and M.K. Veldhuis, vol. 2: 482. Avi Publishing, Westport.Google Scholar
  39. Mansell, R.L. and E.W. Weiler. 1980a. Radioimmunoassay for the determination of limonin in Citrus. Phytochem. 19: 1403–1407.Google Scholar
  40. Mansell, R.L. and E.W. Weiler. 1980b. Immunological tests for the evaluation of citrus quality, in Citrus nutrition and quality, ed. by S. Nagy and J.A. Attaway. ACS Symposium Series 143: 341–359.CrossRefGoogle Scholar
  41. McLaughlin, M.R. and O.W. Barnett. 1979. The influence of plant sap and antigen buffer additives in the enzyme-immunoassay of two plant viruses. Phytopath. 69: 1038.Google Scholar
  42. Mertens, R., Eberle, J., Arnscheidt, A., Ledebur, A. and E.W. Weiler. 1985. Monoclonal antibodies to plant growth regulators. II. Indole-3-acetic acid. Planta 166: 389–393.CrossRefGoogle Scholar
  43. Morgan, M.R.A., McNerney, R., Matthew, J.A., Coxon, D.T. and H.W.S. Chan. 1983. An enzyme-linked immunosorbent assay for total glycoalkaloids in potato tubers. J. Sci. Food Agric. 34: 593–598.CrossRefGoogle Scholar
  44. Nagatani, A., Yamamoto, K.T., Furuya, M., Fukumoto, T. and A. Yamashita. 1984. Production and characterization of monoclonal antibodies which distinguish different surface strutures of pea (Pisum sativum cv. Alaska) phytochrome. Plant & Cell Physiol. 25: 1059–1068.Google Scholar
  45. Narayanan, K.R., Somers, D.A., Kleinhofs, A. and R.L. Warner. 1983. Nature of cytochrome c reductase in nitrate reductase-deficient mutants in barley. Mol. Gen. Genet. 190: 222–226.CrossRefGoogle Scholar
  46. Perrot-Rechenmann, C., Jacquot, J.P., Gadal, P., Weeder, N.F., Cseke, C. and B.B. Buchanan. 1983. Localization of NADP-malate dehydrogenase of corn leaves by immunological methods. Plant Sci. Lett. 30: 219–226.CrossRefGoogle Scholar
  47. Pratt, L.H. 1978. Molecular properties of phytochrome. Photochem. Photobiol. 27: 81–105.CrossRefGoogle Scholar
  48. Rennard, S.I., Church, R.L., Rohrbach, D.H., Shupp, D.E., Abe, S., Hewitt, A.T., Murray, J.C. and G.R. Martin. 1981. Localization of the human fibronectin (FN) gene on chromosome 8 by a specific enzyme immunoassay. Biochem. Genet. 19: 551–566.PubMedCrossRefGoogle Scholar
  49. Robins, R.J., Morgan, M.R.A., Rhodes, M.J.C. and J.M. Furze. 1984a. An enzyme-linked immunosorbent assay for quassin and closely related metabolites. Anal. Biochem. 136: 145–156.PubMedCrossRefGoogle Scholar
  50. Robins, R.J., Morgan, M.R.A., Rhodes, M.J.C. and J.M. Firze. 1984b. Determination of quassin in picogram quantities by an enzyme-linked immunosorbent assay. Phytochem. 23: 1119–1123.CrossRefGoogle Scholar
  51. Rouseff, R.L. 1980. Flavonoids and citrus quality. in Citrus nutrition and quality, ed. by S. Nagy and J.A. Attaway. ACS Symposium Series 143: 83–108.CrossRefGoogle Scholar
  52. Saji, H., A. Nagatani, K.T. Yamamoto, M. Furuya, T. Fukumoto and A. Yamashita. 1984. Cross-reactivity of monoclonal antibodies against rye and pea phytochrome with phytochromes extracted from eight different plant species. Plant Sci. Lett. 37: 57–61.CrossRefGoogle Scholar
  53. Sandberg, G., Ljung, K. and P. Alm. 1985. Precision and accuracy of radioimmunoassay in the analysis of endogenous 3-indoleacetic acid from needles of Scots pine. Phytochem. 24: 1439–1442.CrossRefGoogle Scholar
  54. Saunders, M.J., Cordonnier, M.M., Palevitz, B.A. and L.H. Pratt. 1983. Immunofluorescence visualization of phytochrome in Pisum sativum L. epicotyls using monoclonal antibodies. Planta 159: 545–553.CrossRefGoogle Scholar
  55. Schneider, H.A.W. and W. Leidgens. 1981. An evolutionary tree based on monoclonal antibody-recognized surface features of a plastid enzyme (5-aminolevulinate dehydratase). Z. Naturforsch. 36: 44–50.Google Scholar
  56. Shimazaki, Y., M.-M. Cordonnier and L.H. Pratt. 1983. Phytochrome quantitation in crude extracts of Avena by enzyme-linked immunosorbent assay with monoclonal antibodies. Planta 159: 534–544.CrossRefGoogle Scholar
  57. Spector, S. 1971. Quantitative determination of morphine in serum by radioimmunoassay. J. Pharmacol. Exp. Ther. 178: 253–258.PubMedGoogle Scholar
  58. Thomas, B., Crook, N.E. and S.E. Penn. 1984a. An enzyme-linked immunosorbent assay for phytochrome. Physiol. Plant. 60: 409–415.CrossRefGoogle Scholar
  59. Thomas, B., Penn, S.E., Butcher, G.W. and G. Galfre. 1984b. Discrimination between the red-and far-red-absorbing forms of phytochrome from Avena sativa L. by monoclonal antibodies. Planta 160: 382–384.CrossRefGoogle Scholar
  60. van Weemen, B.K. and A.H.W.M. Schuurs. 1971. Immunoassay using antigen-enzyme conjugates. FEBS Lett. 15: 232–236.PubMedCrossRefGoogle Scholar
  61. Vernooy-Gerritsen, M., Bos, A.L.M., Veldink, G.A. and J.F.G. Vliegenthart. 1983. Affinity chromatography of antibodies directed against soybean lipoxygenase-1 and-2 and an enzyme-linked immunosorbent assay (ELISA) for antibodies and lipoxygenases. Biochim. Biophys. Acta 748: 148–152.PubMedCrossRefGoogle Scholar
  62. Vora, S. 1985. Monoclonal antibodies in enzyme research: present and potential applications. Anal. Biochem. 144: 307–318.PubMedCrossRefGoogle Scholar
  63. Vreeland, V., Slomich, M. and W.M. Laetsch. 1984. Monoclonal antibodies as molecular probes for cell wall antigens of the brown alga, Fucus. Planta 162: 506–517.CrossRefGoogle Scholar
  64. Vreeland, V., Larsen, B. and W.M. Laetsch. 1982. Monoclonal antibodies to Fucus cell wall antigens: localization and specificity patterns. J. Cell Biol. 95: 127a.CrossRefGoogle Scholar
  65. Weigel, U., Horn, W. and B. Hock. 1984. Endogenous auxin levels in terminal stem cuttings of Chrysanthemum morifolium during adventitious rooting. Physiol. Plant. 61: 422–428.CrossRefGoogle Scholar
  66. Weiler, E.W. 1986. Personal communication.Google Scholar
  67. Weiler, E.W. 1984. Immunoassay of plant growth regulators. in Annual Review of Plant Physiology ed. by W.R. Briggs, R.L. Jones and V. Walbot. Annual Reviews (Palo Alto) pp. 85-95.Google Scholar
  68. Weiler, E.W. 1983. Immunoassay of plant consituents. Biochem. Soc. Trans. 11: 485–495.PubMedGoogle Scholar
  69. Weiler, E.W. 1982a. An enzyme immunoassay for cis-(+)-abscisic acid. Physiol. Plant. 54: 510–514.CrossRefGoogle Scholar
  70. Weiler, E.W. 1982b. Plant hormone immunoassay. Physiol. Plant. 54: 230–234.CrossRefGoogle Scholar
  71. Weiler, E.W. 1980. Radioimmunoassay for the differential and direct analysis of free and conjugated abscisic acid in plant abstracts. Planta 148: 262–272.CrossRefGoogle Scholar
  72. Weiler, E.W., Eberle, J., Mertens, R., Atzorn, R., Feyerabend, M., Jourdan, P.S., Arnscheidt, A. and U. Wieczorek. 1986. Antisera-and monoclonal antibody-based immunoassays of plant hormones. in Immunology in Plant Science ed. by T.L. Wang. Cambridge University Press, pp 27-58.Google Scholar
  73. Weiler, E.W., Jourdan P.S. and R.L. Mansell. 1984. Peroxidase-linked, solid-phase enzyme immunoassay for the determination of picomole levels of limonin. Plant Sci. Lett. 35: 159–167.CrossRefGoogle Scholar
  74. Weiler, E.W., Jourdan, P.S. and W. Conrad. 1981. Levels of indole-3-acetic acid in intact and decapitated coleoptiles as determined by a specific and highly sensitive solid-phase enzyme immunoassay. Planta 153: 561–571.CrossRefGoogle Scholar
  75. Weiler, E.W. and R.L. Mansell. 1980. Radioimmunoassay of limonin using a tritiated tracer. J. Agric. Food Chem. 28: 543–545.CrossRefGoogle Scholar
  76. Weiler, E.W. and M.H. Zenk. 1976. Radioimmunoassay for the determination of digoxin and related compounds in Digitalis lanata. Phytochem. 15: 1537–1545.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Richard L. Mansell
    • 1
  • Cecilia A. McIntosh
    • 1
  1. 1.Biology DepartmentUniversity of South FloridaTampaUSA

Personalised recommendations