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Pharmaceutical Research

, Volume 28, Issue 8, pp 1884–1894 | Cite as

Aggregation Stability of a Monoclonal Antibody During Downstream Processing

  • Paolo Arosio
  • Giuliano Barolo
  • Thomas Müller-Späth
  • Hua Wu
  • Massimo Morbidelli
Research Paper

ABSTRACT

Purpose

To study the effect of several operative parameters, particularly pH and salt concentration, on the stability and aggregation kinetics of IgG solutions under the conditions typically encountered in downstream processing.

Methods

The time evolution of the aggregates is analyzed by a combination of dynamic light scattering, size exclusion chromatography (SEC) and field flow fractionation (FFF). Secondary structure changes are monitored by circular dichroism.

Results

For the given antibody, it is found that at pH lower than 4.0 addition of salt induces a reversible aggregation to oligomers accompanied by an increase in the content of the β-sheet structure. The aggregation rate increases monotonically with the salt concentration. Both the SEC and FFF techniques are successfully applied to obtain the oligomer distributions, and their results are consistent. The modified Lumry-Eyring kinetic model can well describe the time evolutions of the oligomers.

Conclusions

For the given antibody, low pH and presence of salt induce conformational changes that are responsible for the reversible aggregation, but in the investigated conditions only small soluble oligomers are formed and oligomer sizes larger than trimer are negligible. For this reason, no accompanied macroscopic changes can be observed.

KEY WORDS

aggregation antibody FFF oligomer distribution SEC 

Notes

ACKNOWLEDGMENTS

Financial support of the Swiss National Science Foundation (Grant No. 200020-126487/1) is gratefully acknowledged. We thank also Merck Serono (Vevey, Switzerland) for supplying the monoclonal antibody and Dr. Silvia Campioni and Prof. Riek (ETH Zurich) for allowing and assisting CD measurements in their labs.

Supplementary material

11095_2011_416_MOESM1_ESM.doc (896 kb)
ESM 1 (DOC 896 kb)

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Paolo Arosio
    • 1
  • Giuliano Barolo
    • 1
  • Thomas Müller-Späth
    • 1
  • Hua Wu
    • 1
  • Massimo Morbidelli
    • 1
  1. 1.Department of Chemistry and Applied Biosciences Institute for Chemical and BioengineeringETH ZurichZurichSwitzerland

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