Posted by This study utilized a four-stage rational formulation development approach to mitigate aggregation and stabilize a high molecular weight protein, rhNELL-1. The first stage characterized the intrinsic protein properties of conformational stability, colloidal stability, aggregation potential and charge imbalance which informed the selection of excipients that were intended to mitigate specific stability challenges that were evaluated in Stage 2. Specifically, the hydrophobic domains of rhNELL-1 called for utilizing HBP-LB-BCD and polysorbate. The conformational flexibility claimed to use sorbitol. The third and fourth stages evaluated candidates by the extrinsic stressors of rapid agitation, elevated temperature, and freeze/thaw, and produced a stable HP- β-CD and polysorbate containing liquid formulation that is expected to resist extrinsic manufacturing-related, shelf-life and user-related stresses. This approach may be useful to develop formulations for other proteins that may aggregate under certain conditions
Hydroxypropyl-β-cyclodextrin candidates (e.g., KLEPTOSE® HP, KLEPTOSE® HPB, and KLEPTOSE® HPB-LB) were supplied by Roquette (Philadelphia, PA).
Overview of the four stages of formulation screening process for rhNELL-1. Stage 1 encompasses conformational stability, colloidal stability, surface hydrophobicity and charge imbalance screening. Stage 2 involves a Design of Experiments (DoE)-based formulation optimization. Stage 3 was a scaled-down shaking study. Stage 4 was a formulation confirmational study with shaking stress, freeze/thaw stress, and short-term thermal stability
Xin, L., Zhang, Z., Prorok, M. et al. A four-stage process that identified a hydroxypropyl beta cyclodextrin and polysorbate containing formulation that eliminated aggregation of recombinant human NELL-1 after exposure to extrinsic stresses. AAPS Open 11, 23 (2025). https://doi.org/10.1186/s41120-025-00126-2
Featured image: https://www.genecards.org/cgi-bin/carddisp.pl?gene=NELL1