Hostname: page-component-8448b6f56d-qsmjn Total loading time: 0 Render date: 2024-04-25T00:15:41.178Z Has data issue: false hasContentIssue false
  • English
  • Français

A monoclonal antibody against Echinococcus multilocularis Em2 antigen

Published online by Cambridge University Press:  06 April 2009

P. Deplazes  and
B. Gottstein
P. Deplazes
Affiliation:
Institute of Parasitology, University of Zürich, Winterthurerstrasse 266a, CH-8057 Zürich, Switzerland
B. Gottstein
Affiliation:
Institute of Parasitology, University of Zürich, Winterthurerstrasse 266a, CH-8057 Zürich, Switzerland
Get access Rights & Permissions [Opens in a new window]

Extract

A monoclonal antibody (MAb G11) species-specific to the Em2 antigen of Echinococcus multilocularis was generated for (i) further biological characterization of the Em2 antigen, (ii) easy affinity-purification of Em2 antigen for immunodiagnostic and immunological investigations and (iii) development of a sandwich-ELISA for the detection of Em2 antigen in diagnostic samples and thus species-specific identification of E. multilocularis metacestode material. The MAb G11 was used in an antibody sandwich-ELISA to detect soluble Em2 antigen with a methodical sensitivity of 80 ng E. multilocularis antigen/ml of solution. MAb G11 specifically detected Em2 antigen in all of 15 E. multilocularis-isolates originating from various geographical areas and in none of other helminth isolates (e.g. Echinococcus granulosus, E. vogeli, and others). Further biological analysis by FITC-labelled MAb G11 demonstrated unique binding activity to the laminated layer of the metacestode. Also, oncospheres were binding FITC-labelled MAb G11 on an outer layer synthesized during cultivation in vitro for 13 days after hatching. Application of the MAb G11 antibody sandwich-ELISA for investigation of solubilized oncospheres confirmed the in vitro synthesis of Em2 antigen by oncospheres on day 13 p.i. Adult stages (somatic antigens) and freshly hatched oncospheres were always MAb G11 negative. Solid-phase MAb G11 was used for purification of the corresponding Em2 antigen by affinity chromatography. A preliminary serological evaluation of the Em2(G11) antigen by ELISA revealed identical immunodiagnostic characteristics, compared to Em2 obtained by classical means, thus suggesting the presented method for future isolation of large-scale Em2 antigen.

Keywords

Echinococcus multilocularisEm2 antigenmonoclonal antibodyaffinity chromatographyEm2-sandwich-ELISA
Type
Research Article
Information
Parasitology , Volume 103 , Issue 1 , August 1991 , pp. 41 - 49
Copyright
Copyright © Cambridge University Press 1991

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Aggarwal, A., Merritt, J. W. & Nash, T. E. (1989). Cysteine-rich variant surface proteins of Giardia lamblia. Molecular and Biochemical Parasitology 23, 39–48.CrossRefGoogle Scholar
Baumann, D. & Gottstein, B. (1987). A double-antibody sandwich ELISA for the detection of Entamoeba histolytica antigen in stool samples of humans. Tropical Medicine and Parasitology 38, 81–5.Google ScholarPubMed
Dao, L. (1985). An improved method of antigen detection on nitrocellulose: in situ staining of alkaline phosphatase conjugated antibody. Journal of Immunological Methods 82, 225–31.Google ScholarPubMed
De St. Groth, F. S. & Scheidegger, D. (1980). Production of monoclonal antibodies: strategy and tactics. Journal of Immunological Methods 35, 1–21.CrossRefGoogle ScholarPubMed
Deplazes, P. & Eckert, J. (1988). Massengewinnung und Lagerung von Taenia hydatigena-Eiern sowie Isolierung lebensfähiger Onkosphären. Schweizer Archiv für Tierheilkunde 130, 307–20.Google Scholar
Eckert, J. & Pohlenz, J. (1976). Zur Wirkung von Mebendazol auf Metazestoden von Mesocestoides corti und Echinococcus multilocularis. Tropical Medicine and Parasitology 27, 247–62.Google ScholarPubMed
Engvall, E. & Perlmann, P. (1972). Enzyme-linked immunosorbent assay, ELISA III. Journal of Immunology 109, 120–35.Google Scholar
Fleischmann, J. B., Pain, R. H. & Porter, R. R. (1962). Reduction of γ-globulin. Archives of Biochemistry and Biophysics 1 (Suppl.), S174–S180.Google Scholar
Gottstein, B. (1985). Purification and characterization of a specific antigen from Echinococcus multilocularis. Parasite Immunology 7, 201–12.CrossRefGoogle ScholarPubMed
Gottstein, B., Eckert, J. & Fey, H. (1983). Serological differentiation between Echinococcus granulosus and E. multilocularis infections in man. Parasitology Research 69, 347–56.Google Scholar
Gottstein, B., Schantz, P. M. & Wilson, J. F. (1985). Serological screening for Echinococcus multilocularis infections with ELISA. Lancet 1, 1097–8.CrossRefGoogle ScholarPubMed
Gottstein, B., Schantz, P. M., Todorov, T., Saimot, A. G. & Jacquier, P. (1986). An international study on the serological differential diagnosis of human cystic and alveolar echinococcosis. Bulletin of the World Health Organization 64, 101–5.Google Scholar
Gottstein, B., Lengeler, C., Bachmann, P., Hagemann, P., Kocher, P., Brossard, M., Witassek, F. & Eckert, J. (1987). Sero-epidemiological survey for alveolar echinococcosis (by Em2-ELISA) of blood donors in an endemic area of Switzerland. Transactions of the Royal Society of Tropical Medicine and Hygiene 81, 960–4.CrossRefGoogle Scholar
Gottstein, B., Tschudi, K., Eckert, J. & Ammann, R. (1989). Em2-ELISA for the follow-up of alveolar echinococcosis after complete surgical resection of liver lesions. Transactions of the Royal Society of Tropical Medicine and Hygiene 83, 389–93.CrossRefGoogle ScholarPubMed
Gottstein, B., Deplazes, P. J., Eckert, J., Müller, E., Schott, E., Helle, O., Boujon, P., Wolff, K., Wandeler, A., Schwiete, U. & Moegle, H. (1991). Serological (Em2-ELISA) and parasitological examinations of fox populations for Echinococcus multilocularis infections. Journal of Veterinary Medicine, Series B (in the Press).CrossRefGoogle ScholarPubMed
Lanier, A. P., Trujillo, D. E., Schantz, P. M., Wilson, J. F., Gottstein, B. & McMahon, B. J. (1987). Comparison of serologic tests for the diagnosis and follow-up of alveolar hydatid disease. American Journal of Tropical Medicine and Hygiene 37, 609–15.CrossRefGoogle ScholarPubMed
Müller, N., Gottstein, B., Vogel, M., Flury, K. & Seebeck, T. (1989). Application of a recombinant Echinococcus multilocularis antigen in an ELISA for diagnosis of human alveolar echinococcosis. Molecular and Biochemical Parasitology 36, 151–60.CrossRefGoogle Scholar
Rausch, R. L, Wilson, J. F., Schantz, P. M. & McMahon, B. J. (1987). Spontaneous death of Echinococcus multilocularis: Cases diagnosed serologically (by Em2-ELISA) and clinical significance. American Journal of Tropical Medicine and Hygiene 36, 576–85.CrossRefGoogle ScholarPubMed
Schantz, P. M. & Gottstein, B. (1986). Echinococcosis (Hydatidosis). In Immunodiagnosis of Parasitic Diseases, vol. 1 (ed. Walls, K. W. & Schantz, P. M), pp. 69–107. Orlando: Academic Press.Google Scholar