Effect of Arginine on Pre-nucleus Stage of Interferon Beta-1b Aggregation

Abstract

Abstract. Understanding the mechanism of aggregation of a therapeutic protein would not only ease the manufacturing processing but could also lead to a more stable finished product. Aggregation of recombinant interferon (IFNβ-1b) was studied by heating, oxidizing, or seeding of unformulated monomeric solution. The formation of aggregates was monitored by dynamic light scattering (DLS) and UV spectroscopy. The autocatalytic monomer loss model was used to fit the data on aggregation rates. The influence of pre-nucleation on aggregation step was demonstrated by inducing the liquid samples containing a monomer form of folded IFNβ-1b by heat and also an oxidizing agent. Results tend to suggest that the nucleus includes a single protein molecule which has been probably deformed. Seeding tests showed that aggregation of IFNβ-1b was probably initiated when 1.0% (w/w) of monomers converted to nucleus form. Chemiluminescence spectroscopy analysis of the sample indicated the generation of 3.0 μM of hydrogen peroxide (H 2 O 2 ) during nucleation stage of IFNβ-1b aggregation. Arginine with a concentration of 200 mM was sufficient to suppress aggregation of IFNβ-1b by decreasing the rate of prenucleation step. We proposed the formation of pre-nucleus structures prior to nucleation as the mechanism of aggregation of IFNβ-1b. Furthermore, we have showed the positive anti-aggregation effect of arginine on pre-nucleation step.

Review of the current state of protein aggregation inhibition from a materials chemistry perspective: special focus on polymeric materials

“…Proteins such as bevacizumab may follow the traditional Arrhenius relationship within a narrow temperature range between 25 1C and 50 1C; 116 however, most proteins do not behave this way for a wide range of temperatures. 117,118 Therefore, the rate of aggregation might be different at 25 1C than at 40 1C 70 due to the change in protein aggregate solubility, 119 various aggregation mechanisms, 120 conformational stability, 121 and the reversibility of protein aggregation 122 at different temperatures. Thus, prescribed storage temperatures for proteins based on accelerated stability data, 121 calculated using a modified Arrhenius equation 121 and the extended Lumry-Eyring (ELE) aggregation model should be followed.…”

Cognitive simulation along with neural adaptation explain effects of suggestions: a novel theoretical framework

“…In other words, regardless of baseline cognitive capabilities, to some extent, all participants can exert top-down control over perception. For example, in two previous studies with healthy participants, all of them showed top-down attenuation of neural activity, regardless of their performance in other tasks ( Lopresti-Goodman et al, 2013 ; Fazeli et al, 2014 ).…”

Inhibiting Protein Aggregation Using Cellulose Nanocrystal in MALDI-TOF MS Analysis: Improving the Sensitivity and Repeatability of Intact Protein in Pueraria

“…Single protein in the sample solution associates with another protein to form oligomer as the initial aggregation event and eventually lead to formation of higher multimers. The smallest aggregate is dimer, and the upper limit of size depends on protein properties and environmental conditions. , Usually, a larger molecular size can induce a low desorption/ionization efficiency and a decreased sensitivity in MALDI-TOF MS analysis. , Protein aggregation may be induced by undesirable solution conditions, the presence of a destabilizer/organic solvent, long-term storage, or under a single or several stress conditions, which may be observed in all stages of sample preparation and analysis. − Several interventions, including chemical modification or mutation, lyophilization, and use of stabilizers, can be taken to inhibit protein aggregation. Site-specific mutations have proved to be effective in controlling protein aggregation, however, this approach is complex and may potentially affect protein integrity. − Alternatively, protein stabilizers control the protein aggregation tendency mainly by reducing protein interactions and enhancing the conformational stability, such as amino acid derivatives, polysorbates, polyols, and sugars. − It is also noted that protein stabilizers applied to one stress condition may be unsuitable to another due to potential differences in aggregation mechanisms.…”