Preparation and characterization of a monoclonal antibody against mannoprotein of Candida albicans.
BALB/c mice were immunized via injection with whole cell of Candida albicans serotype A. The spleens were fused with myeloma cells of SP2/0 origin. A mannoprotein-reactive monoclonal antibody (MAb) was selected and characterized by ELISA technique. This MAb reacted with strains of Candida such as C. albicans, C. tropicalis, and C. albicans of the Persian Type Culture Collection (PTCC). However, our antibody did not react with other Candida species such as C. parapsilosis, C. glabrata, C. stellatoidae, C. lusitania, C. krusei, and S. cervisiae. These antibodies also did not recognize extracts of other fungal species such as Aspergillus fumigatus and Aspergillus flavus, and bacterial strains such as Staphylococcus aureus and Pseudomonas aeruginosa. Polyclonal antibody produced in this study could not differentiate the above species and was reactive towards all fungal species mentioned above except bacterial strains of S. aureus and P. aeruginosa. Western blot analysis of ligand affinity-purified mannoproteins of C. albicans wall protein using this MAb showed reactivity toward a single protein band in the region of 55-65 kDa molecular weight. The same antibody, when examined with unpurified C. albicans extract, reacted with a broad band in the region of 55-105 kDa, which we concluded was due to a possible different glycosylation pattern of mannoprotein in crude extract in which the higher molecular weight protein was eliminated by ligand-binding affinity purification.
Farahnejad
Z
Z
Department of Medical Mycology, School of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.
Rasaee
M J
MJ
Moghadam
M Frozandeh
MF
Paknejad
M
M
Kashanian
S
S
Rajabi
M
M
D000818:Animals; D000908:Antibodies, Fungal / Q000096:biosynthesis / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D000946:Antigens, Fungal / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D002176:Candida albicans / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000276:immunology*; D002473:Cell Wall / Q000737:chemistry / Q000276:immunology; D004586:Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005656:Fungal Proteins / Q000276:immunology / Q000302:isolation & purification; D006825:Hybridomas; D007114:Immunization; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D012703:Serotyping
D000908:Antibodies, Fungal; D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; C025363:mannoproteins