Development and in vitro characterization of a humanized scFv against fungal infections.
Tomas Di MambroTania VanzoliniMarzia BianchiRita CrinelliBarbara CanonicoFilippo TasiniMichele MenottaMauro MagnaniPublished in: PloS one (2022)
The resistance and the birth of new intrinsic and multidrug-resistant pathogenic species like C. auris is creating great concern in the antifungal world. Given the limited drug arsenal and the lack of effectiveness of the available compounds, there is an urgent need for innovative approaches. The murine mAb 2G8 was humanized and engineered in silico to develop a single-chain fragment variable (hscFv) antibody against β-1,3-glucans which was then expressed in E. coli. Among the recombinant proteins developed, a soluble candidate with high stability and affinity was obtained. This selected protein is VL-linker-VH oriented, and it is characterized by the presence of two ubiquitin monomers at the N-terminus and a His tag at the C-terminus. This construct, Ub2-hscFv-His, guaranteed stability, solubility, efficient purification and satisfactory recovery of the recombinant product. HscFv can bind β-1,3-glucans both as coated antigens and on C. auris and C. albicans cells similarly to its murine parental and showed long stability and retention of binding ability when stored at 4°, -20° and -80° C. Furthermore, it was efficient in enhancing the antifungal activity of drugs caspofungin and amphotericin B against C. auris. The use of biological drugs as antifungals is limited; here we present a promising hscFv which has the potential to be useful in combination with currently available antifungal drugs.
Keyphrases
- candida albicans
- multidrug resistant
- monoclonal antibody
- induced apoptosis
- randomized controlled trial
- systematic review
- drug induced
- drug resistant
- cell cycle arrest
- small molecule
- acinetobacter baumannii
- gram negative
- emergency department
- dendritic cells
- oxidative stress
- room temperature
- molecular docking
- gestational age
- pseudomonas aeruginosa
- klebsiella pneumoniae
- transcription factor
- cystic fibrosis
- cell proliferation