Interactions of formulation excipients with proteins in solution and in the dried state

Ohtake, Satoshi; Kita, Yoshiko; Arakawa, Tsutomu

doi:10.1016/j.addr.2011.06.011

Cited by 322 publications

(276 citation statements)

References 248 publications

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“…20 Current approaches to mitigate chemical and physical instabilities require the addition of buffers and excipients to mAb formulations. 21 Regarding protein adsorption and aggregation as discussed above, the industry generally uses non-ionic surfactants such as Polysorbate 20 and 80 (Tween Ò 20 and 80) to control shake-induced aggregation as would be experienced during drug product transportation, for instance. In general, polysorbates compete with the protein for an interface and adsorb to exposed hydrophobic patches on the protein surface.…”

Section: Introductionmentioning

confidence: 99%

Antibody adsorption on the surface of water studied by neutron reflection

Smith

Holman

et al. 2017

mAbs

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Surface and interfacial adsorption of antibody molecules could cause structural unfolding and desorbed molecules could trigger solution aggregation, resulting in the compromise of physical stability. Although antibody adsorption is important and its relevance to many mechanistic processes has been proposed, few techniques can offer direct structural information about antibody adsorption under different conditions. The main aim of this study was to demonstrate the power of neutron reflection to unravel the amount and structural conformation of the adsorbed antibody layers at the air/water interface with and without surfactant, using a monoclonal antibody 'COE-3 0 as the model. By selecting isotopic contrasts from different ratios of H 2 O and D 2 O, the adsorbed amount, thickness and extent of the immersion of the antibody layer could be determined unambiguously. Upon mixing with the commonly-used non-ionic surfactant Polysorbate 80 (Tween 80), the surfactant in the mixed layer could be distinguished from antibody by using both hydrogenated and deuterated surfactants. Neutron reflection measurements from the co-adsorbed layers in null reflecting water revealed that, although the surfactant started to remove antibody from the surface at 1/100 critical micelle concentration (CMC) of the surfactant, complete removal was not achieved until above 1/10 CMC. The neutron study also revealed that antibody molecules retained their globular structure when either adsorbed by themselves or co-adsorbed with the surfactant under the conditions studied.

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Section: Introductionmentioning

confidence: 99%

Antibody adsorption on the surface of water studied by neutron reflection

Smith

Holman

et al. 2017

mAbs

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“…the mAb tested ("mAb-1") showed highest surface loading to silica at pH 7.4 (~12 mg/m Excipients are often included in protein formulations to stabilize the monomeric form and reduce aggregation and surface adsorption. 9 Many examples can be drawn on to illustrate the utility of polysorbates in protein bioprocessing, formulation and delivery. In general, polysorbates compete with the protein for an interface and adsorb to exposed hydrophobic patches on the protein surface.…”

Section: Introductionmentioning

confidence: 99%

Adsorption behavior of a human monoclonal antibody at hydrophilic and hydrophobic surfaces

Couston

Skoda

Uddin

et al. 2013

mAbs

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“…Apparent variation in the chemical and physical properties of amino acid excipients (e.g., pH), as well as their electrostatic interaction with protein molecules, may explain the larger variation in their mixing states than those with sugars and sugar alcohols. 21) In addition, the size and structure of the proteins may also affect thermodynamic and kinetic factors determining the mixing states. Further clarification of factors determining the phase behavior may contribute to formulation developments.…”

Section: Resultsmentioning

confidence: 99%

“…Formation of physically stable crystalline cake makes glycine (Gly) a popular bulking agent, while maintenance of amorphous state should be preferable for pH buffering agents (e.g., L-histidine (L-His)) and the stabilizers. [19][20][21] Mixing state of the protein with excipient molecules in the amorphous freeze-concentrate and in dried solids is another physical factor determining quality of multi-component freeze-dried protein pharmaceuticals. 19,22,23) Ice growth during the freezing segment of the lyophilization process significantly concentrates the multiple solutes into a narrow space.…”

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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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Interactions of formulation excipients with proteins in solution and in the dried state

Cited by 322 publications

References 248 publications

Antibody adsorption on the surface of water studied by neutron reflection

Antibody adsorption on the surface of water studied by neutron reflection

Adsorption behavior of a human monoclonal antibody at hydrophilic and hydrophobic surfaces

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

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