Oligoethylene glycols prevent thermal aggregation of α‐chymotrypsin in a temperature‐dependent manner: Implications for design guidelines

Tomita, Shunsuke; Tanabe, Yumiko; Shiraki, Kentaro

doi:10.1002/btpr.1762

Cited by 3 publications

(2 citation statements)

References 46 publications

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“…This is because the heatinduced inactivation mainly results from the chemical modifications of proteins. It is interesting to note that the suppressive effect of Arg on heat-induced aggregation of αchymotrypsin retains constant when temperature different from 65°C to 85°C, while that of a small poly(ethylene glycol) of tetraethylene glycol (TEG) increases with increasing temperature [91]. This results from the increase in hydrophobic interaction between TEG and unfolded protein with increasing temperature.…”

Section: Difference Between Heat-induced Degradation and Refolding-inmentioning

confidence: 99%

Small Amine Molecules: Solvent Design Toward Facile Improvement of Protein Stability Against Aggregation and Inactivation

Shiraki

Tomita²,

Inoue³

2015

CPB

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Proteins are prone to inactivation in aqueous solutions because chemical modification and aggregation usually occur, particularly at high temperature. This review focuses on the recent advance in practical application with amine compounds that prevent the heat-induced inactivation and aggregation of proteins. Coexistence of amine solutes, typically diamines, polyamines, amino acid esters, and amidated amino acids decreases the heat-induced inactivation rate of proteins by one order of magnitude compared with that in the absence of additives under low concentrations of proteins at physiological pH. The amine compounds mainly suppress chemical modification, typically the β-elimination of disulfide bond and deamidation of asparagine side chain, thereby preventing heat-induced inactivation of proteins. Polyamines do not improve the refolding yield of proteins, owing to decrease in the solubility of unfolded proteins. In contrast, arginine is the most versatile additive for various situations, such as refolding of recombinant proteins, solubilized water-insoluble compounds, and prevention of nonspecific binding to solid surfaces; however, it is not always effective for preventing heat-induced aggregation. Amine compounds will be a key to prevent protein inactivation in solution additives.

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Section: Difference Between Heat-induced Degradation and Refolding-inmentioning

confidence: 99%

Small Amine Molecules: Solvent Design Toward Facile Improvement of Protein Stability Against Aggregation and Inactivation

Shiraki

Tomita²,

Inoue³

2015

CPB

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“…The third point also relates to irreversible aggregate formation between thermally unfolded molecules. For example, oligoethylene glycols (PEG units) suppress the irreversible aggregation of α-chymotrypsin at higher temperatures [105]. Thus, the repulsive force between complexes of cationic enzyme and cationic PEAMA-g-PEG is higher than that between enzyme molecules alone.…”

Section: Stabilization Of Protein By Pegylated Polymermentioning

confidence: 99%

Wrap-and-Strip Technology of Protein–Polyelectrolyte Complex for Biomedical Application

Shiraki¹,

Kurinomaru²,

Tomita³

2016

CMC

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Two Elution Mechanisms of MEP Chromatography

Arakawa

Tokunaga

Maruyama

et al. 2018

CPPS

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MEP (mercapto-ethyl-pyridine) HyperCel is one of the hydrophobic charge induction chromatography (HCIC) resins. Under normal operation, proteins are bound to the MEP resin at neutral pH, at which MEP is not charged, mostly via hydrophobic interaction. MEP has a pyridine group, whose pK is 4.8, and hence is positively charged at acidic pH range. Based on the binding mechanism (i.e., hydrophobic interaction) and the induced positive charge at acidic pH, there may be two ways to elute the bound proteins. One way is to bring the pH down to protonate both MEP resin and the bound protein, leading to charge repulsion and thereby elution. Another way is to use hydrophobic interaction modifiers, which are often used in hydrophobic interaction chromatography, to reduce hydrophobic interaction. Here, we summarize such two possible elution approaches.

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Oligoethylene glycols prevent thermal aggregation of α‐chymotrypsin in a temperature‐dependent manner: Implications for design guidelines

Cited by 3 publications

References 46 publications

Small Amine Molecules: Solvent Design Toward Facile Improvement of Protein Stability Against Aggregation and Inactivation

Small Amine Molecules: Solvent Design Toward Facile Improvement of Protein Stability Against Aggregation and Inactivation

Wrap-and-Strip Technology of Protein–Polyelectrolyte Complex for Biomedical Application

Two Elution Mechanisms of MEP Chromatography

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