Mechanisms of protein stabilization in the solid state

Chang, Liuquan; Pikal, Michael J.

doi:10.1002/jps.21825

Cited by 271 publications

(220 citation statements)

References 122 publications

Supporting

Mentioning

208

Contrasting

Order By: Relevance

“…This enzyme stabilization is the result of the occurrence of different interactions between the enzyme and the adjuvants used. However, the mechanisms of stabilization in the solid state are still not completely understood (Chang and Pikal, 2009). From the enzymatic activity results for spray dried lipase, it was concluded that the enzyme activity of dried lipase using β-cyclodextrin was higher than with the other adjuvants.…”

Section: Spray Drying Stabilization Of Purified Lipasementioning

confidence: 99%

Characterization and spray drying of lipase produced by the endophytic fungus Cercospora kikuchii

Costa-Silva

Souza

Oliveira

et al. 2014

Braz. J. Chem. Eng.

View full text Add to dashboard Cite

-A lipase from the endophytic fungus Cercospora kikuchii was purified, biochemically characterized and the effects of spray drying on stabilization of the purified enzyme were studied. The lipase was purified 9.31-fold with recovery of 26.6% and specific activity of 223.6 U/mg. The optimum pH and temperature were 4.6 and 35 °C, respectively, while the Vmax was 10.28 µmol/min.mg -1 protein and Km 0.0324 mM. All the metal ions tested enhanced the enzyme activity. The lipase retained almost 100% activity in the presence of strong oxidants and was also resistant to Triton X, Tween 80 and 20 and SDS, as well as to proteases. The purified lipase was spray dried and kept until 85.2% of enzymatic activity. At least 70% of the enzymatic activity was maintained for spray dried purified lipase during the storage period. The lipase produced by Cercospora kikuchii has properties useful for industrial application and showed adequate stabilization and retention of its enzymatic activity after spray drying.

show abstract

Section: Spray Drying Stabilization Of Purified Lipasementioning

confidence: 99%

Characterization and spray drying of lipase produced by the endophytic fungus Cercospora kikuchii

Costa-Silva

Souza

Oliveira

et al. 2014

Braz. J. Chem. Eng.

View full text Add to dashboard Cite

show abstract

“…Proteins and excipients are required to be concentrated into the non-crystalline solute-mixture phase to achieve the stabilizing effects. 13) Among the three segments comprising the lyophilization process, (freezing, primary drying, and secondary drying), optimization of the product temperature during the time-consuming ice-subliming primary drying segment, by controlling the shelf temperature and chamber pressure is inevitable for achieving appropriate formulation quality and process efficiency. 1,[14][15][16] A higher product temperature during the primary drying allows faster ice sublimation, 17) whereas decreased viscosity of the non-crystalline concentrated solute phase increases the risk of solid structure change (collapse) that starts from the sublimation interface.…”

mentioning

confidence: 99%

Component Crystallization and Physical Collapse during Freeze-Drying of <small>L</small>-Arginine–Citric Acid Mixtures

Yamaki

Ohdate

Nakadai

et al. 2012

CHEMICAL & PHARMACEUTICAL BULLETIN

View full text Add to dashboard Cite

Component crystallization and physical collapse during freeze-drying of aqueous solutions containing protein-stabilizing L-arginine and citric acid mixtures were studied. Freeze-drying microscopy (FDM) and thermal analysis of the solute-mixture frozen solutions showed collapse onset at temperatures (T c ) approximately 10°C higher than their T g ′s (glass transition temperatures of the maximally freeze-concentrated solute phase). Experimental freeze-drying of these solutions at a low chamber pressure showed the occurrence of physical collapse at shelf temperatures close to or slightly higher than the T c . Slower ice sublimation at higher chamber pressures induced the physical collapse from lower shelf temperatures. The large effect of chamber pressures on the collapse-inducing shelf temperatures confirmed significance of the sublimation-related heat loss on the sublimation interface temperature during the primary drying. Drying of the single-solute L-arginine solution resulted in cake-structure solids composed of its anhydrous crystal. Thermal and powder X-ray diffraction (PXRD) analysis suggested slow crystal nucleation of L-arginine dihydrate in the frozen solutions. Characterization of the frozen solutions and freeze-dried solids should enable rational formulation design and process control of amino acid-containing lyophilized pharmaceuticals.Key words freeze-drying; crystallization; glass; lyophilization; collapse; thermal analysis Clinical applications of protein pharmaceuticals such as therapeutic antibodies are increasing. However, marginal storage stabilities of many proteins in the aqueous solutions emphasize the importance of developing freeze-dried formulations. Many lyophilized protein formulations contain disaccharides (e.g., sucrose and trehalose) that protect the proteins thermodynamically from structural changes during the freeze-drying process and kinetically from chemical degradation during subsequent storage.1,2) Some basic amino acid (e.g., L-arginine and L-histidine) and multivalent acid (e.g., H 3 PO 4 and citric acid) mixtures are practical alternative stabilizers for protein formulations.3-6) The mixtures protect proteins from conformation-altering dehydration stress during the process. These mixtures form glass-state amorphous solids through networks of hetero-component molecular interactions between the amino and carboxyl groups, and thus improve chemical stability of the dried proteins during storage. 5,7)L-Arginine also increases solubility of various proteins without apparent changes in their conformation. [8][9][10] Freeze-drying is a typical high-energy process, and varying its parameters significantly varies the formulation quality, including the cake appearance, component crystallinity, residual water, and stability of the active pharmaceutical ingredients (APIs).11,12) Failure of a biopharmaceutical lyophilization batch would lead to the loss of the therapeutic protein often more expensive than small molecular APIs. Freezing of aqueous solutions concentrates most solutes to a s...

show abstract

“…The most common approach towards stabilising protein drugs is to remove the water from the formulation (13,14), often in the presence of specific excipients which prevent protein unfolding due to dehydration stress (15,16). Water tends to decrease the shelf life of proteins by increasing the molecular mobility of the protein, and acts as a reactant in several degradation pathways (14).…”

Section: Introductionmentioning

confidence: 99%

Design of Experiments-Based Monitoring of Critical Quality Attributes for the Spray-Drying Process of Insulin by NIR Spectroscopy

Maltesen

Weert

2012

AAPS PharmSciTech

View full text Add to dashboard Cite

Abstract. Moisture content and aerodynamic particle size are critical quality attributes for spray-dried protein formulations. In this study, spray-dried insulin powders intended for pulmonary delivery were produced applying design of experiments methodology. Near infrared spectroscopy (NIR) in combination with preprocessing and multivariate analysis in the form of partial least squares projections to latent structures (PLS) were used to correlate the spectral data with moisture content and aerodynamic particle size measured by a time of flight principle. PLS models predicting the moisture content were based on the chemical information of the water molecules in the NIR spectrum. Models yielded prediction errors (RMSEP) between 0.39% and 0.48% with thermal gravimetric analysis used as reference method. The PLS models predicting the aerodynamic particle size were based on baseline offset in the NIR spectra and yielded prediction errors between 0.27 and 0.48 μm. The morphology of the spray-dried particles had a significant impact on the predictive ability of the models. Good predictive models could be obtained for spherical particles with a calibration error (RMSECV) of 0.22 μm, whereas wrinkled particles resulted in much less robust models with a Q 2 of 0.69. Based on the results in this study, NIR is a suitable tool for process analysis of the spray-drying process and for control of moisture content and particle size, in particular for smooth and spherical particles.

show abstract

Mechanisms of protein stabilization in the solid state

Cited by 271 publications

References 122 publications

Characterization and spray drying of lipase produced by the endophytic fungus Cercospora kikuchii

Characterization and spray drying of lipase produced by the endophytic fungus Cercospora kikuchii

Component Crystallization and Physical Collapse during Freeze-Drying of <small>L</small>-Arginine–Citric Acid Mixtures

Design of Experiments-Based Monitoring of Critical Quality Attributes for the Spray-Drying Process of Insulin by NIR Spectroscopy

Contact Info

Product

Resources

About