Partial characterization of a species-specific stachybotrys chartarum antigen.
Schmechel-D; Hettick-J; Simpson-J; Beezhold-D
J Allergy Clin Immunol 2005 Feb; 115(2)(Suppl 1):S165
The presence of Stachybotrys in water-damaged homes and businesses has received a great deal of attention beacuse this mold produces several potent mycotoxins, but the association between Stachybotrys and building related disease is controversial. We have developed a species-specific spore-specific monoclonal antibody (MAb) 9B4 that could be used for monitoring environmental contamination. The purpose of this work was to characterize the antigen recognized by the MAb 9B4. Stachybotrys chartarum (29-51-05) was grown on MEA medium until sporulation occurred and the spores collected by rolling glass beads over the dry cultures. Spores were eluted from the beads and extracted with PBS-T. The extracts were analyzed for the presence of the antigen. By SDS-PAGE and Western blot analysis we observed an antigen of approximately 20 kD. MALDI-TOF analysis determined the MW to be 19.65 kD. The antigen was found to be trypsin sensitive but insensitive to periodate treatment or enzymatic deglycosylation indicating it to be a protein. 2-D Western Blot analysis indicated 2 isoforms of the antigen with a pI between 4.5-5. Amino acid compositional analysis of affinity-purified antigen indicated the protein is rich in glycine and serine and the sequence for one tryptic decapeptide has been obtained. Our data indicates that the antigen recognized by 9B4 is a unique 19.6 kD non-glycoylated protein with a pI of approximately 5. Because this antigen carries species-specific information it could serve as an important marker to quantify for environmental exposure assessment.
Antigens; Molds; Bacteria; Diseases; Bacterial-disease; Environmental-contamination; Monitoring-systems; Environmental-exposure; Exposure-assessment; Microorganisms; Mass-spectrometry
Abstract; Conference/Symposia Proceedings
Research Tools and Approaches: Exposure Assessment Methods
The Journal of Allergy and Clinical Immunology