PREPARATION OF ANTI-PROTEIN AND ANTIMANNAN ANTISERA AGAINST FUNGAL CELL-WALL BY AFFINITY-CHROMATOGRAPHY

A novel and easy chromatographic method has been developed for the iso lation of anti-protein and anti-mannan antisera from a population of p olyclonal antibodies obtained against Candida albicans and Yarrowia li polytica cell wall manno-proteins. The technique is based on the immob ilization of mannan (to be used as immunoadsorbent) by Affi-Prep H(z) resin after the oxidation of neighboring hydroxyl groups of the polysa ccharide with sodium periodate, For Y. lipolytica polyclonal antiserum , a single chromatographic step using the homologous mannan was suffic ient to obtain an antiprotein antibody preparation free of antimannan antibodies. For C. albicans, three chromatographic processes using hom ologous and heterologous mannan were needed to obtain a satisfactory a ntiprotein antiserum. The potential application of the anti-protein an tiserum obtained has been demonstrated by indirect immunofluorescence assays of whole cells and electrophoretic analysis of wall proteins in C. albicans and Saccharomyces cerevisiae. (C) 1994 Academic Press, In c.