Production and characterization of three monoclonal antibodies to Candida albicans proteins

Abstract

Three monoclonal antibodies, designated A2C7, C2C7, and F19, were produced which recognize proteins from Candida albicans. All are of the immunoglobulin GI heavy chain and kappa light chain class. A2C7 and C2C7 immunoprecipitated three proteins contained in a partially purified fraction (region A) of a mycelial cytoplasmic extract of C. albicans. The apparent molecular weights of these proteins are 120,000 (120K) to 135K, 44K to 52K, and 35K to 38K. Monoclonal antibody F19 was reactive with proteins of 42K, 43K, and 50K in immunoblotting experiments. F19 was also able to form a precipitin band in agarose gel with protein(s) contained in region A. Limited proteolytic digestion of the three proteins immunoprecipitated by A2C7 and C2C7 demonstrated that both monoclonal antibodies recognized the same three Candida proteins and that there exists a significant degree of relatedness in primary structure among the three proteins. Proteins with apparent molecular weights of 120K to 135K, 44K to 52K, and 35K to 38K that were immunoprecipitated by sera from two patients with invasive candidiasis and by the serum from a rabbit immunized against a 48K (44K to 52K) Candida protein were also analyzed by limited proteolysis. Patterns of peptide fragments generated by enzymatic digestion of these proteins showed that the proteins recognized by the monoclonal antibodies are the same proteins recognized by antibodies in the sera of patients during an invasive Candida infection and by antibodies in the serum of the immune rabbit.

Enhanced IgE response to Candida albicans in postoperative invasive candidiasis

“…In immunoblotting an immunodominant 47 kDa protein antigen of C. albicans to IgG antibodies has been recognized as enolase [34][35][36][37]. Another important antigen with a molecular weight of 47 kDa has been identified as a splitdown product of a fungal heat shock protein 90 [38].…”

Evaluation of latex reagents for rapid identification of Candida albicans and C. krusei colonies

“…The agglutination test is a simple procedure which has been applied for many years to the study of yeast surface antigens (39). Several monoclonal antibodies have been proposed for the identification of C. albicans (4,18,23,28,33,39); however, the expression of their antigenic epitopes at the surface of the yeast cell was shown to be extremely variable, and they were unable to agglutinate C. albicans cells with sufficient sensitivity and specificity. Thus, the Bichro-latex albicans test is the first immunological reagent useful for the rapid, easy to perform, and reliable identification of C. albicans colonies.…”

“…For the rapid identification of yeast colonies, immunological tests with species-specific antisera have been described, which use either polyclonal (38,40) or monoclonal antibodies (4,8,18,23,28,33,39), but these methods either were limited by the lack of specificity of some sera, especially for the species C. albicans and Candida tropicalis (5), or were not commercially available. Recently, new monoclonal antibody-based latex tests have been proposed (Fumouze Diagnostics, Asnières, France) for the rapid identification of C. albicans (34) and C. krusei colonies.…”

Stimulation of human B cells specific for Candida albicans for monoclonal antibody production

“…In view of this, the treatment of systemic candidosis with anti-Candida antibodies is an attractive possibility. There have already been several reports on the production of murine mAbs to various antigenic determinants of Candida such as proteins [24], cell wall components [25][26][27] and nuclear components [28]. As discussed in the INTRODUCTION, the production of a human mAb would be preferable for therapeutic purposes.…”