Targeting Proteins of Aspergillus fumigatus for Monoclonal Antibody Production
AuthorMoschetti, Anthony Steven
AdvisorKozel, Thomas R.
Biochemistry and Molecular Biology
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IA is increasingly prevalent and problematic among patients with immunodeficiency. Early diagnosis of IA can be difficult because the symptoms patients display are not specific to fungal infections, and the diagnostic tests currently in use are unreliable. It is well known that the organisms that cause IA shed protein antigens into body fluids once inside the host. In previous studies, our lab detected PPIase, Eglc, Ecm33, TrxR and CatB in IA mouse serum and urine, using an antigen discovery platform. In our current work, we seek to develop mAbs that specifically target PPIase, Eglc, Ecm33, TrxR and CatB as an effective diagnostic tool for the invasive fungus Aspergillus fumigatus. rPPIase, rEglC and rCatB were purified from bacterial cultures. rPPIase and rCatB were emulsified in TiterMax® Gold adjuvant (CytRx Corporation, Los Angeles, CA) and used to immunize BALB/c mice. The rPPIase emulsion elicited a robust immune response (high titers) which was confirmed via ELISA. The rPPIase immunized mice that had the highest titers were then boosted with recombinant protein in preparation for a splenocyte and myeloma fusion to produce a hybridoma cell line. A mAb was then selected and derived with high specificity and sensitivity for rPPIase. The long term goal is to produce high quality anti-PPIase, anti-Eglc, anti-Ecm33, anti-TrxR and anti-CatB mAbs as candidates for IA diagnostic tests, and possibly as a therapeutic option for IA patients.