Application of isothermal chemical denaturation to early-stage formulation development of fibrinogen

The purpose of this study was to develop fibrinogen formulation using isothermal chemical denaturation (ICD), which is a technique that measures the conformational stability of proteins through structure perturbation by denaturants such as urea. Fibrinogen has been used to treat various congenital and acquired human fibrinogen-related disorders that represent a group of rare conditions. In this study, 10 formulations of fibrinogen were initially prepared and appropriate formulations were screened via automated, high-throughput ICD measurements. The optimal formulation selection process was further performed through osmolality measurements and accelerated stability study at 40°C and 75% relative humidity. The fibrinogen formulation was optimized with 20 mg/ml fibrinogen, 16 mg/ml L-arginine, 10 mg/ml glycine, 1.2 mg/ml tri-sodium citrate dihydrate, and 3.5 mg/ml sodium chloride at pH 7.0. This study demonstrates that ICD-based formulation screening approach is highly applicable to early-stage formulation development of fibrinogen.