Development and Use of a Monoclonal Antibody Specific for the Candida albicans Cell-Surface Protein Hwp1

Abstract

The Candida albicans cell-surface protein Hwp1 functions in adhesion to the host and in biofilm formation. A peptide from the Gln-Pro-rich adhesive domain of Hwp1 was used to raise monoclonal antibody (MAb) 2-E8. MAb 2-E8 specificity for Hwp1 was demonstrated using a hwp1/hwp1 C. albicans isolate and strains that expressed at least one HWP1 allele. Immunofluorescence and atomic force microscopy experiments using MAb 2-E8 confirmed C. albicans germ-tube-specific detection of the Hwp1 protein. MAb 2-E8 also immunolabeled the tips of some Candida dubliniensis germ tubes grown under conditions that maximized HWP1 expression. The phylogeny of HWP1 and closely related genes suggested that the Gln-Pro-rich adhesive domain was unique to C. albicans and C. dubliniensis focusing the utility of MAb 2-E8 on these species. This new reagent can be used to address unanswered questions about Hwp1 and its interactions with other proteins in the context of C. albicans biology and pathogenesis.

Promising immunotherapeutic targets for treating candidiasis

“…Recently, Hoyer and co-workers developed a monoclonal antibody against Hwp1 of C. albicans. The Gln-Pro-rich adhesive domain (CDNPPQPDQPDDN (amino acids 154 to 166 of the protein)) of Hwp1 was utilized to produce monoclonal antibody MAb 2-E8 (Oh et al, 2022). The result showed that 2-E8 can bind to Hwp1 specifically.…”

“…The result showed that 2-E8 can bind to Hwp1 specifically. MAb 2-E8 labeled the germ tube of the wild-type strain but not the hwp1D/D strain (Oh et al, 2022). Furthermore, assays were performed to assess the blocking potential of MAb2-E8 on adhesive interactions.…”

“…As a control, Anti-Als1 MAb1-B2, which was previously shown to inhibit C. albicans adhesion to biliary epithelial cells (BECs), was included (Coleman et al, 2010). Results indicated that MAb2-E8 at a 20 µg/mL concentration effectively blocked adhesion, like Anti-Als1 (Oh et al, 2022). Furthermore, Rosario-Colon et al found that Candida Hwp1-specific monoclonal antibodies 6H1 could protect mice against invasive C. auris infection by significantly enhancing survival and reducing fungal burdens (Rosario-Colon et al, 2021).…”

Analytical characterization of host‐cell‐protein‐rich aggregates in monoclonal antibody solutions

“…DDA data from all native digest samples and the standard digest of SPF 2 were used to construct a consolidated spectral library in Skyline as described previously (v20.2.0.343; MacCoss Lab, University of Washington). 51,56,57 Triplicate SWATH data for each sample were processed together using the Skyline commandline interface, and peaks were selected and integrated automatically using the mProphet algorithm based on a target decoy approach. 58 The consolidated spectral library was used in the analysis of all samples except the PAVIN mAb SEC fraction, which contained too few HCP peptides for mProphet automatic peak picking.…”