Structure-based discovery of small molecules improving stability of human broadly-neutralizing anti-HIV antibody 2F5 in plant suspension cells.
Manoj K MandalThales KronenbergerMarie I ZulkaBjörn WindshügelAndreas SchiermeyerPublished in: Biotechnology journal (2022)
The production of biopharmaceuticals in engineered plant-based systems is a promising technology that has proven its suitability for the production of various recombinant glyco-proteins that are currently undergoing clinical trials. However, compared to mammalian cell lines, the productivity of plant-based systems still requires further improvement. A major obstacle is the proteolytic degradation of recombinant target proteins by endogenous plant proteases mainly from the subtilisin family of serine proteases. In the present study, the authors screened for putative small molecule inhibitors for subtilases that are secreted from tobacco BY-2 suspension cells using an in silico approach. The effectiveness of the substances identified in this screen was subsequently tested in degradation assays using the human broadly-neutralizing anti-HIV monoclonal antibody 2F5 (mAb2F5) and spent BY-2 culture medium as a model system. Among 16 putative inhibitors identified by in silico studies, three naphthalene sulfonic acid derivatives showed inhibitory activity in in vitro degradation assays and are similar to or even more effective than phenylmethylsulfonyl fluoride (PMSF), a classical inhibitor of serine proteases, which served as positive control.
Keyphrases
- small molecule
- monoclonal antibody
- induced apoptosis
- high throughput
- endothelial cells
- clinical trial
- antiretroviral therapy
- cell cycle arrest
- hiv positive
- hiv infected
- human immunodeficiency virus
- hepatitis c virus
- hiv testing
- hiv aids
- induced pluripotent stem cells
- drinking water
- molecular docking
- cell wall
- men who have sex with men
- pluripotent stem cells
- endoplasmic reticulum stress
- cell death
- climate change
- oxidative stress
- systematic review
- protein kinase
- cell proliferation
- dengue virus
- cell free
- open label
- single cell
- study protocol
- pi k akt