Figure 3. Mapping of the serospecificities of VL patients against L. donovani antigens.

The proteins of total lysates of L. donovani isolate BHU2 (MHOM/IN/02/BHU2) promastigotes were separated by 2-dimensional gel electrophoresis with a pH gradient of 4.5–7 in the first dimension and a 12.5% SDS-PAGE in the second.

A: Western blot developed with autologous serum.

330 Leishmania antigens could be counted.

B: The corresponding silver-stained gel displaying 1067 protein spots.

68 of 330 antigens could be assigned to protein spots in the silver-stained gel.

Six of the protein spots, indicated by arrows and numbered 1–6, were processed for protein identification by mass spectrometry.

C: Inner section of a SyPro Ruby-stained gel with the same protein material.

The proteins that match antigens detected by the Western blot analysis shown in Panel A are circled.

54 antigenic proteins are thus marked, 14 antigenic proteins are outside the sections shown.

The arrows again indicate the six identified proteins.