title: Aggregation suppression of proteins by arginine during thermal unfolding.
abstract: Arginine has been used to suppress aggregation of proteins during refolding and purification. We have further studied in this paper the aggregation-suppressive effects of arginine on two commercially important proteins, i.e., interleukine-6 (IL-6) and a monoclonal antibody (mAb). These proteins show extensive aggregation in aqueous buffers when subjected to thermal unfolding. Arginine suppresses aggregation concentration-dependently during thermal unfolding. However, this effect was not specific to arginine, as guanidine hydrochloride (GdnHCl) at identical concentrations also was effective. While equally effective in aggregation suppression during thermal unfolding, arginine and GdnHCl differed in their effects on the structure of the native proteins. Arginine showed no apparent adverse effects on the native protein, while GdnHCl induced conformational changes at room temperature, i.e., below the melting temperature. These additives affected the melting temperature of IL-6 as well; arginine increased it concentration-dependently, while GdnHCl increased it at low concentration but decreased at higher concentration. These results clearly demonstrate that arginine suppresses aggregation via different mechanism from that conferred by GdnHCl.
authors.affiliation: Alliance Protein Laboratories, Thousand Oaks, CA 91360, USA. tarakawa2@aol.com
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Arakawa
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Kita
authors.forename: Daisuke
authors.initials: D
authors.lastname: Ejima
authors.forename: Kouhei
authors.initials: K
authors.lastname: Tsumoto
authors.forename: Harumi
authors.initials: H
authors.lastname: Fukada
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001120:Arginine / Q000737:chemistry*; D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D019791:Guanidine / Q000737:chemistry; D006801:Humans; D015850:Interleukin-6 / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D015850:Interleukin-6; D001120:Arginine; D019791:Guanidine