Effects of formulation development methods on the stability of model protein pharmaceuticals embedded in solid lipid matrices

Bioanalytical Workflow for Qualitative and Quantitative Assessment of Hot-Melt Extruded Lysozyme Formulations

“…Typically, standard procedures for the analytical characterization of proteins cover methods such as SDS-PAGE, mass spectrometry, various chromatographic and spectroscopic methods, and functional studies among others. , For our study, to address the FDA guidelines, we aimed to utilize robust and reproducible techniques to evaluate parameters such as purity, identity, quantity, and activity for lysozyme before and after formulation as well as for stored formulated lysozyme samples. For this reason, we investigated two different activity assays, that is a turbidimetric assay and a fluorogenic assay, as well as SDS-PAGE and RP-HPLC analysis for purity determination.…”

“…Therefore, many proteins were selected and tested as models to study the effect of different formulation techniques and their process parameters (such as temperature, applied pressure, or processing time) on protein stability, properties, and activity. 5 Lysozyme, that is, N-acetylmuramide glycanhydrolase or muramidase, is an enzyme that is commonly used for pharmaceutical formulations by different techniques such as freeze drying, 6,7 hot-melt extrusion, 8,9 and numerous other polymer-based techniques as a control. 10−12 Lysozyme cleaves β-1,4-glycosidic bonds between N-acetylmuraminic acid and N-acetylglucosamine.…”

“…Before expanding to large-scale processing, several bench-scale studies are required. Therefore, many proteins were selected and tested as models to study the effect of different formulation techniques and their process parameters (such as temperature, applied pressure, or processing time) on protein stability, properties, and activity …”