Arginine Inhibits Adsorption of Proteins on Polystyrene Surface

Abstract

Nonspecific adsorption of protein on solid surfaces causes a reduction of concentration as well as enzyme inactivation during purification and storage. However, there are no versatile inhibitors of the adsorption between proteins and solid surfaces at low concentrations. Therefore, we examined additives for the prevention of protein adsorption on polystyrene particles (PS particles) as a commonly-used material for vessels such as disposable test tubes and microtubes. A protein solution was mixed with PS particles, and then adsorption of protein was monitored by the concentration and activity of protein in the supernatant after centrifugation. Five different proteins bound to PS particles through electrostatic, hydrophobic, and aromatic interactions, causing a decrease in protein concentration and loss of enzyme activity in the supernatant. Among the additives, including arginine hydrochloride (Arg), lysine hydrochloride, guanidine hydrochloride, NaCl, glycine, and glucose, Arg was most effective in preventing the binding of proteins to PS particles as well as activity loss. Moreover, even after the mixing of protein and PS particles, the addition of Arg caused desorption of the bound protein from PS particles. This study demonstrated a new function of Arg, which expands the potential for application of Arg to proteins.

Arginine prevents thermal aggregation of hen egg white proteins

“…Arg has a guanidinium group on the side chain with the most basic isoelectric point of about pH 10.8. Arg has been applied to solution sciences and industries, e.g., for increasing solubility of poorly soluble compounds, such as drug substances of alkyl gallates (Ariki, Hirano, Arakawa, & Shiraki, 2011;Hirano, Kameda, Arakawa, & Shiraki, 2010), coumarin (Hirano, Arakawa, & Shiraki, 2008), caffeic acid (Hirano, Kameda, Shinozaki, Arakawa, & Shiraki, 2013), and an unfolded protein (Reddy K., Lilie, Rudolph, & Lange, 2005); viscosity control of pharmaceutical proteins (Inoue, Takai, Arakawa, & Shiraki, 2014a, 2014b; adsorption control of proteins onto a solid surface (Hirano, Maruyama, Shiraki, Arakawa, & Kameda, 2014;Shikiya, Tomita, Arakawa, & Shiraki, 2013); improvement of protein refolding (Buchner & Rudolph, 1991;Tsumoto et al, 1998;Umetsu et al, 2003); solubilization of porcine myosin (Takai, Yoshizawa, Ejima, Arakawa, & Shiraki, 2013); and protein crystallization (Ito et al, 2011). In particular, the potential to suppress protein aggregation is one of the major applications of Arg as a solution additive (Das et al, 2007;Golovanov, Hautbergue, Wilson, & Lian, 2004;Sharma, Verma, Singh, Korpole, & Ashish, 2016;Shiraki et al, 2004;Shiraki, Kudou, Fujiwara, Imanaka, & Takagi, 2002).…”

Arginine and lysine reduce the high viscosity of serum albumin solutions for pharmaceutical injection

“…We have developed the application of ArgHCl for suppressing aggregation and adsorption of various proteins during heat treatment and oxidative refolding (16)(17)(18)(19)(20)(21)(22). These effects have been ascribed, at least in part, to the interaction of aromatic groups on arginine with the protein surface (23).…”

A Robust Method for Assaying the Immunoreactive Fraction in Nonequilibrium Systems

“…To evaluate the error introduced by changes in T, three matrices of theoretical B T data are constructed, one with the original B T values and the other two with B T transformed by the Equation (30) for an error ∆T T of ± 1%. This error percentage is chosen to approximate the imprecision of a micropipette with a viscous liquid [28].…”

“…A standard 96-well high-binding PS microplate binds proteins by hydrophobic interactions and π-π stacking [30]. The binding capacity of a high-binding plate is approximately 500 ng mouse IgG/cm 2 and the diameter of the well is 6.4 mm [27].…”