Pharmaceutical Research

, Volume 30, Issue 3, pp 735–750 | Cite as

Liquid Formulations for Stabilizing IgMs During Physical Stress and Long-Term Storage

  • Monika Mueller
  • Maybelle Q. T. Loh
  • Rupert Tscheliessnig
  • Doris H. Y. Tee
  • Eddy Tan
  • Muriel Bardor
  • Alois Jungbauer
Research Paper

ABSTRACT

Purpose

To develop a liquid formulation for IgMs to survive physical stress and storage.

Methods

Stabilizing formulations for 8 monoclonal immunoglobulin (IgMs) were found using differential scanning calorimetry (DSC). In these formulations, the IgMs were subjected to stress and storage and analyzed by size exclusion chromatography and fluorescence activated cell sorting. Structure was analyzed using small-angle X-ray scattering (SAXS).

Results

The highest conformational stability was found near the isoelectric point and further enhanced by addition of sorbitol, sucrose and glycine. For 2 IgMs, the pH optimum for conformational and storage stability did not correspond. Lowering the pH led to the desired storage stability. Optimized formulations prevented aggregation and fragmentation from shear stress, freeze-thaw cycles, accelerated storage and real time storage at 4°C and −20°C for 12 months. Optimized formulations also preserved immunoreactivity for 12 months. SAXS indicated that IgM in stabilizing conditions was closer to the structural IgM model (2RCJ) and less susceptible for aggregation.

Conclusions

A long-term stabilizing formulation for 8 IgMs was found comprising 20% sorbitol and 1 M glycine at pH 5.0–5.5 which may have broad utility for other IgMs. Formulation development using DSC and accelerated storage was evaluated in this study and may be used for other proteins.

KEY WORDS

conformational stability DSC formulation IgM protein aggregation 

Supplementary material

11095_2012_914_MOESM1_ESM.pdf (1 mb)
ESM 1(PDF 1.01 mb)

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Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Monika Mueller
    • 1
  • Maybelle Q. T. Loh
    • 1
  • Rupert Tscheliessnig
    • 2
    • 3
  • Doris H. Y. Tee
    • 1
  • Eddy Tan
    • 1
  • Muriel Bardor
    • 1
  • Alois Jungbauer
    • 3
    • 4
  1. 1.Bioprocessing Technology Institute A*STAR (Agency for Science Technology and Research)SingaporeSingapore
  2. 2.Department of Chemical and Biomolecular EngineeringUniversity of CaliforniaBerkeleyUSA
  3. 3.Austrian Centre of Industrial Biotechnology (ACIB)ViennaAustria
  4. 4.Department of BiotechnologyUniversity of Natural Resources and Life Sciences ViennaViennaAustria

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