Immunoblotting identifies and antigen recognized by anti gp63 in the immune complexes of Indian kala-azar patient sera |
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Authors: | Tapati Sanyal Dilip K Ghosh Dwijen Sarkar |
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Affiliation: | (1) Leishmania Group and Department of Cell Biology, Indian Institute of Chemical Biology, 4, Raja S.C. Mullick Road, Jadavpur, 700 032 Calcutta, India |
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Abstract: | In SDS-PAGE the immune complexes (IC) of kala-azar patient sera showed intense bands at 55 kDa and 20 kDa corresponding to heavy and light chains of immunoglobulins. In immunoblot experiment, kala-azar and normal IC after treatment with patient sera showed multiple bands of which the band at 55 kDa was most prominent in kala-azar IC. It is known that in kala-azar sera antihuman IgG is present, so the heavy band at 55 kDa region may be due to higher amount of IgG and/or other antigen(s) present at that region. Immunoblot experiments of kala-azar IC with anti gp63 also developed a major band at 55 kDa. It suggests that the antigen (55 kDa) and gp63 have common antigenic epitope (s). Normal IC did not react with anti gp63 indicating absence of this antigen in normal IC. Antigenic similarity between the IC antigen (55 kDa) and gp63 indicated that the former antigen may have been processed from gp63. In summary, identification of a parasite antigen (55 kDa) in IC of kala-azar patients sera may be useful in developing a serodiagnostic assay for visceral leishmaniasis. (Mol Cell Biochem130: 11–17, 1994)Abbreviations IC
Immune Complexes
- PEG
Polyethylene Glycol (Mol wt 8000)
- PBS
Phosphate Buffer Saline
- VL
Visceral Leishmaniasis
- AVL
American Visceral Leishmaniasis
- IgG
Immunoglobulin G
- TBS
Tris Buffer Saline
- SDS-PAGE
Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis
- gp63
A leishmanial surface glycoprotein of molecular mass 63,000
- TEMED
N,N,N ,N -Tetramethylethylenediamine |
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Keywords: | immunodiagnosis kala-azar immune complexes anti gp63 Leishmania donovani |
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