FIG. 2.
(A) Purification scheme for MAb 6C5-H4CA antigen isolation. Soluble material released from mechanically broken C. albicans cells was separated by two-dimensional PAGE separation. Coomassie-stained gels were aligned with Western blottings of duplicate gels to identify protein spots reactive with the antibody. The blot was stained with colloidal gold to label protein and was subsequently used for antibody detection. The arrow indicates the protein spot that was recovered for mass spectrometric sequence determination. Molecular masses (in kilodaltons) are shown on the left. The pH gradient of the first dimension is shown on the top. (B) Mass spectrometric analysis of MAb 6C5-H4CA antigen. The DNA sequence of a C. albicans gene product identified by mass spectrometric analysis of the MAb 6C5-H4CA antigen is presented here, with single-letter amino acid symbols of the translated open reading frame underneath. Tryptic fragments identified by mass spectrometry are underlined, and a sequence matching that of the antibody epitope (Fig. 1) is boxed.