Effect of buffer species on the unfolding and the aggregation of humanized IgG

Daisuke Kameoka, Etsuro Masuzaki, Tadashi Ueda, Taiji Imoto

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

The aggregation propensity of humanized antibody after heat treatment is evaluated in the presence of six buffer species. The comparison under equivalent pH showed high aggregation propensity on phosphate and citrate buffer. In contrast, 2-(N-Morpholino) ethane sulfonate (MES), 3-(N-Morpholino) propane sulfonate (MOPS), acetate and imidazole buffer showed lower aggregation propensity than the above two buffers. Meanwhile, unfolding temperature evaluated by differential scanning calorimetry measurement was not altered among these buffer species. The light scattering analysis suggested that heat-denatured intermediate was aggregated slightly on MES and acetate buffer. Therefore, it was found that the different aggregation propensity among buffer species was caused from the aggregation propensity of heat-denatured intermediate rather than the unfolding temperature. Furthermore, it was revealed that the aggregation dependency on buffer species is accounted for by the specific molecular interaction between buffer and IgG, rather than the ionic strength. On the contrary, on the analyses of unfolding and aggregation propensity by molecular dissection of IgG into Fab and Fc fragments, aggregation propensity of Fc fragment on MES, acetate and phosphate buffer was almost the same as whole IgG. From the above results, it was suggested that the specific interaction between buffer molecule and Fc domain of IgG was involved in the aggregation propensity of heat-denatured IgG.

Original languageEnglish
Pages (from-to)383-391
Number of pages9
JournalJournal of Biochemistry
Volume142
Issue number3
DOIs
Publication statusPublished - Sep 1 2007

Fingerprint

Buffers
Agglomeration
Immunoglobulin G
Morpholinos
Hot Temperature
Immunoglobulin Fc Fragments
Acetates
Phosphates
Antibodies, Monoclonal, Humanized
Dissection
Propane
Immunoglobulin Fab Fragments
Temperature
Molecular interactions
Differential Scanning Calorimetry
Ionic strength
Citric Acid
Light scattering
Osmolar Concentration
Differential scanning calorimetry

All Science Journal Classification (ASJC) codes

  • Medicine(all)
  • Biochemistry
  • Molecular Biology

Cite this

Effect of buffer species on the unfolding and the aggregation of humanized IgG. / Kameoka, Daisuke; Masuzaki, Etsuro; Ueda, Tadashi; Imoto, Taiji.

In: Journal of Biochemistry, Vol. 142, No. 3, 01.09.2007, p. 383-391.

Research output: Contribution to journalArticle

Kameoka, Daisuke ; Masuzaki, Etsuro ; Ueda, Tadashi ; Imoto, Taiji. / Effect of buffer species on the unfolding and the aggregation of humanized IgG. In: Journal of Biochemistry. 2007 ; Vol. 142, No. 3. pp. 383-391.
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