Monoclonal antibodies 3C3, 6D4, 7D11, 9G6, 24D11, 27C10, 27E2, and 29E5.
Hybridoma 2011 Feb; 30(1):103
Antigen Used for Immunization: Stachybotrys chlorohalonata (ATCC 201863) cytolytic antigens (cScp) were semi-purified from tryptic soy broth culture supernatants as previously described. Method of Immunization: Mice were immunized intraperitoneally at biweekly intervals. Mice were primed with 50 mg of cScp emulsified in equal volumes of TiterMax (TiterMax USA, Norcross, GA). The antigen concentration was reduced by half for each of five subsequent booster immunizations. A final boost of 50 mu g was given 3 weeks after the sixth immunization, and mice were sacrificed 3 days later for hybridoma production. Parental Cell Line Used for Fusion: SP2/0-AG14 myleoma cells (ATCC# CRL-1581). Selection and Cloning Procedure: Positive clones were identified using 1 mu g/mL of cScp in an indirect ELISA. Heavy and Light Chains of Immunoglobulin: Heavy chain - mu. Light chain - not determined. Specificity: Hybridoma tissue culture supernatants (CSN) were screened using an indirect ELISA as previously described. The specificity of MAbs was tested against hyphal extracts of seven S. chartarum isolates, eight isolates of other Stachybotrys species, as well as 39 related and non-related fungi commonly found in indoor environments. All extracts were tested in a capture ELISA using rabbit polyclonal antibodies as solidphase capture reagent. Specific binding activity was documented using Western blot analysis against mycelial extracts derived from Stachybotrys chlorohalonata and using fluorescent halogen immunostaining with various fungal species. Specific Antigen Identified: Exoantigens produced by Stachybotrys chlorohalonata. Availability: Tissue culture supernatant: Yes; Ascitic fluid: No; Hybridoma cells: Yes.
Allergic-reactions; Antibody-response; Antigens; Biological-effects; Biological-factors; Cytotoxic-effects; Fungi; Immune-reaction; Immune-system; Immunology; Laboratory-animals; Laboratory-testing; Mycology; Mycotoxins; Toxic-effects; Microorganisms
Donald H. Beezhold, Ph.D., Allergy and Clinical Immunology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, 1095 Willowdale Road, M/S L-4020, Morgantown, WV 265