Spectroscopic evaluation of the stabilization of humanized monoclonal antibodies in amino acid formulations

Tian, Fei; Middaugh, C. Russell; Offerdahl, Tom; Munson, Eric J.; Sane, Samir U.; Rytting, J. Howard

doi:10.1016/j.ijpharm.2006.10.037

Cited by 81 publications

(49 citation statements)

References 62 publications

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“…Actually, m-DOPA lowers the ablation threshold of the target thus allowing using lower laser pulse energy, as indicated in table 2. Furthermore, m-DOPA acts as an excipient to protect lipase from denaturation induced during plume expansion by drying [12]. Pentane is added to induce lid opening and is responsible for the microemulsion formation.…”

Section: Resultsmentioning

confidence: 99%

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Frozen Microemulsions for MAPLE Immobilization of Lipase

Califano

Bloisi

Perretta

et al. 2017

Preprint

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MAPLE (matrix assisted pulsed laser evaporation) depositions of Candida Rugosa lipase were carried out from ice matrices whose composition is optimized in order to minimize conformational damage of the protein, which strongly influences its catalytic activity. To induce lid opening and to protect lipase during the MAPLE process, pentane and m-DOPA amino acid were added to the liquid matrix giving a target formed by a frozen water-lipase-pentane microemulsion. FTIR and AFM were used to investigate the structure of MAPLE deposited lipase films. The ability of MAPLE films to promote transesterification was determined by thin layer chromatography. It was shown that m-DOPA has influence on the aggregation but not on the unfolding of lipase induced by MAPLE, while the microemulsion formed by the addition of pentane to the target composition is effective in protecting lipase during the MAPLE process. MAPLE deposited lipases showed a modified specificity.

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Section: Resultsmentioning

confidence: 99%

“…In this work, m-DOPA is added to reduce the risk of photochemical damages acting as a laser radiation absorber and to protect lipase against unfolding/aggregation that can occur during drying in plume expansion [12], whereas pentane is added to induce lid opening.…”

Section: Introductionmentioning

confidence: 99%

Frozen Microemulsions for MAPLE Immobilization of Lipase

Califano

Bloisi

Perretta

et al. 2017

Preprint

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“…In addition, the lower effective concentration of monosodium citrate compared to sucrose suggested the contribution of electrostatic (ion-ion, ion-dipole) interactions between the salt anion (hydroxycarboxylate ion) and basic amino acid residues on protein surfaces for the structural stabilization. 17,36,37) The high T g of the dried solids indicated mixing of the protein and salts required for the interaction. The topic of protein-stabilizing molecular interactions in the dried states, including the effect of differently ionized functional groups, will require further study for elucidation.…”

Section: Discussionmentioning

confidence: 99%

Stabilization of Protein Structure in Freeze-Dried Amorphous Organic Acid Buffer Salts

Izutsu¹,

Kadoya

Yomota³

et al. 2009

Chem. Pharm. Bull.

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The development of protein pharmaceuticals requires rational formulation design to ensure appropriate storage stability, because the degradation of such pharmaceuticals through various chemical and physical pathways not only reduces their therapeutic effects but also increases the risk of product immunogenicity. [1][2][3][4][5] Freeze-drying is a popular method of conferring long-term stability of therapeutic proteins that is not achievable in aqueous solutions. Removal of the surrounding water molecules during the freeze-drying process, however, often perturbs the protein structure, leading to irreversible aggregation in the reconstituted solutions. The structurally altered protein molecules are also prone to chemical degradation during storage.1) Maintaining the protein conformation by process and ingredient (e.g., stabilizer, pH buffer, isotonic agents) optimization thus improves both the physical and chemical stability of protein formulations.Choosing the solution pH and buffer system appropriate to a particular protein is a simple but significant element in the formulation design because the chemical and physical integrity of proteins in the aqueous solutions and freeze-dried solids depend largely on the pH. 6) Some buffer components also favorably or adversely affect the protein stability through direct interactions and/or through changing the local environments in the dried state. For example, freezing of certain buffer systems (e.g., sodium phosphate) often induces crystallization of a component salt and resulting shift of the local pH surrounding the proteins.7-11) Freeze-drying from some buffer systems (e.g., L-histidine, citrate, or Tris) often leads to higher activity retention of proteins (e.g., coagulation factor VIII, recombinant human interleukin-1 receptor antagonist) relative to those from other buffers.12-15) Conformation of the proteins lyophilized in these buffer systems is of particular interest.Reported properties of some carboxylic acid salts, including stabilization of native protein conformation in aqueous solutions (e.g., antithrombin III) 16,17) and their propensity to form glass-state amorphous solids upon lyophilization, 18) suggest their ability to protect protein conformation against dehydration stress through mechanisms similar to disaccharides. Non-reducing disaccharides (e.g., sucrose, trehalose) are popular stabilizers in solution and freeze-dried protein formulations. Various saccharides and polyols thermodynamically favor native protein structures over denatured states in aqueous solutions by a "preferential exclusion" mechanism. 19) Sucrose and trehalose protect proteins by substituting surrounding water molecules through hydrogen bonds during the freeze-drying process. 4,[20][21][22] Limited molecular mobility in glass-state lyophilized disaccharide solids also protects embedded proteins from chemical degradation (e.g., deamidation) during storage. 23)The present study assesses the physical properties and protein-stabilizing effects of carboxylic acid buffer systems (e.g.,...

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“…5,8,[12][13][14][15] Physical states (e.g., crystallinity, crystal polymorph) of the components are important factors that determine chemical and conformational stability of proteins during the freeze-drying process and subsequent storage. [16][17][18] The stabilizing effects of disaccharides are attributed to protection of protein conformation by the substitution of surrounding water molecules and by holding protein molecules in lower molecular mobility glassstate amorphous solids.…”

mentioning

confidence: 99%

“…[5][6][7][8][9][10][11] Application of the amino acid excipients that protect proteins through particular mechanisms not achievable by saccharides (e.g., aggregation-reducing effect of L-arginine (L-Arg) in aqueous solution) would increase formulation strategies in lyophilization of marginally stable proteins. 5,8,[12][13][14][15] Physical states (e.g., crystallinity, crystal polymorph) of the components are important factors that determine chemical and conformational stability of proteins during the freeze-drying process and subsequent storage. [16][17][18] The stabilizing effects of disaccharides are attributed to protection of protein conformation by the substitution of surrounding water molecules and by holding protein molecules in lower molecular mobility glassstate amorphous solids.…”

mentioning

confidence: 99%

Amorphous–Amorphous Phase Separation of Freeze-Concentrated Protein and Amino Acid Excipients for Lyophilized Formulations

Izutsu¹,

Yoshida²,

Shibata³

et al. 2016

CHEMICAL & PHARMACEUTICAL BULLETIN

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The objective of this study was to elucidate the mixing state of proteins and amino acid excipients concentrated in the amorphous non-ice region of frozen solutions. Thermal analysis of frozen aqueous solutions was performed in heating scans before and after a heat treatment. Frozen aqueous solutions containing a protein (e.g., recombinant human albumin, gelatin) or a polysaccharide (dextran) and an amino acid excipient (e.g., L-arginine, L-arginine hydrochloride, L-arginine monophosphate, sodium L-glutamate) at varied mass ratios showed single or double T g ′ (glass transition temperature of maximally freeze-concentrated solutes). Some mixture frozen solutions rich in the polymers maintained the single T g ′ of the freeze-concentrated amorphous solute-mixture phase. In contrast, amino acid-rich mixture frozen solutions revealed two T g ′s that suggested transition of concentrated non-crystalline solute-mixture phase and excipient-dominant phase. Post-freeze heat treatment induced splitting of the T g ′ in some intermediate mass ratio mixture solutions. The mixing state of proteins and amino acids varied depending on their structure, salt types, mass ratio, composition of co-solutes (e.g., NaCl) and thermal history. Information on the varied mixing states should be valuable for the rational use of amino acid excipients in lyophilized protein pharmaceuticals.Key words amorphous; freeze drying; thermal analysis; protein formulation; stabilization; mixing Freeze-drying is a popular method to formulate therapeutic proteins that are insufficiently stable in aqueous solutions during storage.1) Several freeze-dried therapeutic protein formulations contain multiple excipients, including stabilizers, bulking agents, pH buffer agents, and tonicity modifiers besides active pharmaceutical ingredient (API).2-4) Sucrose and trehalose are popular stabilizers that protect proteins from dehydration-induced irreversible structural changes during the process and from chemical degradation during storage. Certain amino acids and their salts are potent stabilizers in the freeze-drying of proteins, frozen storage of liposomes, and spray drying of vaccines.5-11) Application of the amino acid excipients that protect proteins through particular mechanisms not achievable by saccharides (e.g., aggregation-reducing effect of L-arginine (L-Arg) in aqueous solution) would increase formulation strategies in lyophilization of marginally stable proteins. 5,8,[12][13][14][15] Physical states (e.g., crystallinity, crystal polymorph) of the components are important factors that determine chemical and conformational stability of proteins during the freeze-drying process and subsequent storage. [16][17][18] The stabilizing effects of disaccharides are attributed to protection of protein conformation by the substitution of surrounding water molecules and by holding protein molecules in lower molecular mobility glassstate amorphous solids. Crystallization of some sugar alcohols (e.g., mannitol) during the freezing segment of lyophilization deprives the...

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Spectroscopic evaluation of the stabilization of humanized monoclonal antibodies in amino acid formulations

Cited by 81 publications

References 62 publications

Frozen Microemulsions for MAPLE Immobilization of Lipase

Frozen Microemulsions for MAPLE Immobilization of Lipase

Stabilization of Protein Structure in Freeze-Dried Amorphous Organic Acid Buffer Salts

Amorphous–Amorphous Phase Separation of Freeze-Concentrated Protein and Amino Acid Excipients for Lyophilized Formulations

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