Enhanced protein aggregation suppressor activity of N-acetyl-L-arginine for agitation-induced aggregation with silicone oil and its impact on innate immune responses

Nam Ah Kim, Ga Yeon Noh, Shavron Hada, Kyung Jun Na, Hee Jung Yoon, Ki Woong Park, Young Min Park, Seong Hoon Jeong

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Previously, N-acetyl-L-arginine (NALA) suppressed the aggregation of intravenous immunoglobulins (IVIG) more effectively and with a minimum decrease in transition temperature (Tm) than arginine monohydrochloride. In this study, we performed a comparative study with etanercept (commercial product: Enbrel®), where 25 mM arginine monohydrochloride (arginine) was added to the prefilled syringe. The biophysical properties were investigated using differential scanning calorimetry (DSC), dynamic light scattering (DLS), size-exclusion chromatography (SEC), and flow-imaging microscopy (FI). NALA retained the transition temperature of etanercept better than arginine, where arginine significantly reduced the Tm by increasing its concentration. End-over-end rotation was applied to each formulation for 5 days to accelerate protein aggregation and subvisible particle formation. Higher monomeric content was retained with NALA with a decrease in particle level. Higher aggregation onset temperature (Tagg) was detected for etanercept with NALA than arginine. The results of this comparative study were consistent with previous study, suggesting that NALA could be a better excipient for liquid protein formulations. Agitated IVIG and etanercept were injected into C57BL/6J female mice to observe immunogenic response after 24 h. In the presence of silicone oil, NALA dramatically reduced IL-1 expression, implying that decreased aggregation was related to reduced immunogenicity of both etanercept and IVIG.

Original languageEnglish
Pages (from-to)42-51
Number of pages10
JournalInternational Journal of Biological Macromolecules
Volume216
DOIs
StatePublished - 1 Sep 2022

Keywords

  • Acetyl arginine
  • Arginine monohydrochloride
  • Immunogenicity
  • Protein aggregation
  • Protein formulation
  • Subvisible particle

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