The study was conducted with the prime objective of isolating an antigen from the crude preparation of whole promastigotes (Leishmania donovani) with a view to future exploitation in serodiagnosis and production of monoclonal antibodies. Soluble antigen, prepared from promastigotes isolated from actively growing cultures, was fractionated by gel filtration chromatography over a column of Sephadex G200. Three peaks of proteins could be recovered. The antigenic reactivity of different fractions was checked against sera of kala azar patients by immunoelectrophoresis (IEP), rocket immunoelectrophoresis (RIEP) and enzyme-linked immunosorbent assay (ELISA). The first peak was found to be highly reactive in comparison with other peaks. SDS-PAGE and western blot analysis of this antigen revealed a major antigen of promastigotes in the vicinity of 65 to 66 kDa, while a weakly reactive triplet could also be detected in the low molecular weight region.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Enzyme-Linked Immunosorbent Assay
Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis
Brain Heart Infusion
Hanks' Balanced Salt Solution
Diamino Benzidine Tetrahydrochloride
Arora SK, Datta U, Sehgal S (1985) Use of excretory facor of L. mexicana for serodiagnosis of kala azar: A preliminary report. Ind J Med Res 81:580–583
Clinton BB, Palezuk MC, Stauber LA (1972) L. donovani: Partial characterization of some flagellate cytoplasmic immunogens. J Immunol 108:1570–1577
Colomer-Gould V, Glavas Quintao L, Keithly J, Nogucira N (1985) A common major surface antigen on amastigotes and promastigotes of leishmania species. J Exp Med 163:902–916
Etges RJ, Bouvier J, Haffman R, Bordier C (1985) Evidence that the major surface proteins of three leishmania species are structurally related. Mol Biochem Parasitol 14:141–149
Ghosh AC, Row DS (1976) Antigenic components of the protozoan parasites L. enrietti inducing humoral and CMI responses of infected guinae pigs. Immunochem 14:459–465
Graber P, Burtin P (1964) Immunoelectrophoresis analysis. Elsevier Publishing Company, Amsterdam
Laemmli UK (1970) Changes of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680–685
Laurell CB (1972) Electrophoretic and electrochemical analysis of proteins. Scand J Clin Invest 290:124
Lemesre JL, Rizvi FS, Afchain D, Sadigursky M, Capron A, Santoro F (1985) Subspecies specific surface antigens of promastigotes of the L. donovani complex. Infect Immun 50:136–141
Lowry OH, Rosebrough NJ, Fase AL, Randall RJ (1951) Protein measurement by Folin's phenol reagent. J Biol Chem 193:265–275
Misle JA, Marquez ME, Hernandez AC (1985) Surface component of L. braziliensis. Parasitenkd 71:419–428
Peters W, Prasad LSN (1983) Kala azar in India, its importance as an issue in public health. In: Proceedings of Indo-UK Workshop on Leishmaniasis pp 5–9
Preston PM, Dumonde DC (1976) Immunology of clinical and experimental leishmaniasis. In: Cohen S, Sadun EH (eds) Immunology of parasite infections. Blackwell Sc Publ, Oxford, pp 167
Ramasamy R, Kar SK, Jamnadas H (1983) Cross reacting surface antigens on leishmania promastigotes. Int J Parasitol 13:337–341
Sinha R, Arora SK, Sehgal S (1988) A sensitive ELISA for detection of circulating antigens in kala azar patients. Ind J Med Res 88:138–140
Towbin H, Strachellin T, Gordon J (1979) Blotting of proteins from SDS-PAGE and their analysis. Proc Natl Acad Sci (USA) 76:4350–4354
Voller A, Bartlett A, Bidwell DE (1976) Enzyme immunoassays for parasitic diseases. Trans Roy Soc Trop Med Hyg 70:98–106
About this article
Cite this article
Arora, S.K., Sehgal, S. Identification of major antigens of Leishmania donovani using kala azar sera. Med Microbiol Immunol 178, 81–88 (1989). https://doi.org/10.1007/BF00203303
- Western Blot
- Blot Analysis
- Western Blot Analysis
- Immunosorbent Assay
- Prime Objective