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The Role of Polysorbate 80 and HPβCD at the Air-Water Interface of IgG Solutions

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Abstract

Purpose

To test the hypothesis of surface displacement as the underlying mechanism for IgG stabilization by polysorbates and HPβCD against surface-induced aggregation.

Methods

Adsorption/desorption-kinetics of IgG-polysorbate 80/-HPβCD were monitored. Maximum bubble pressure method was used for processes within seconds from surface formation. Profile analysis tensiometry was applied over long periods and to assess surface rheologic properties. Additionally, the kinetics of adsorption, desorption and surface displacement was followed by a double-capillary setup of the profile analysis tensiometer, allowing drop bulk exchange.

Results

Weak surface activity for HPβCD vs. much higher surface activity for polysorbate 80 was shown. Protein-displacement when exceeding a polysorbate 80 concentration close to the CMC and a lack of protein displacement for HPβCD was observed. The drop bulk exchange experiments show IgG displacement by polysorbate 80 independent of the adsorption order. In contrast, HPβCD coexists with IgG at the air-water interface when the surface layer is built from a mixed IgG-HPβCD-solution. Incorporation of HPβCD in a preformed IgG-surface-layer does not occur.

Conclusions

The results confirm surface displacement as the stabilization mechanism of polysorbate 80, but refute the frequently held opinion, that HPβCD stabilizes proteins against aggregation at the air-water interface in a manner comparable to non-ionic surfactants.

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Abbreviations

BSA:

bovine serum albumin

CDs:

cyclodextrins

CMC:

critical micellar concentration

HPβCD:

hydroxypropyl-β-cyclodextrin

IgG:

immunoglobulin G

mAbs:

monoclonal antibodies

MBPM:

maximum bubble pressure method

Rh-GCSF:

recombinant human granulocyte colony-stimulating factor

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Acknowledgments and Disclosures

The authors thank the German National Academic Foundation (Studienstiftung des Deutschen Volkes) for financial support. Thanks are also expressed to Roche Diagnostics GmbH for material support.

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Authors and Affiliations

  1. Dept. of Pharmacy, Pharmaceutical Tech. & Biopharmaceutics, Ludwig-Maximilians-University, Butenandtstrasse 5-13, 81377, Munich, Germany

    Tim Serno, Elisabeth Härtl, Ahmed Besheer & Gerhard Winter

  2. Max-Planck-Institut für Kolloid- und Grenzflächenforschung, Am Mühlenberg 1, 14424, Potsdam, Germany

    Reinhard Miller

  3. Novartis Pharma AG, 4002, Basel, Switzerland

    Tim Serno

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  1. Tim Serno
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  2. Elisabeth Härtl
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Correspondence to Tim Serno.

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Serno, T., Härtl, E., Besheer, A. et al. The Role of Polysorbate 80 and HPβCD at the Air-Water Interface of IgG Solutions. Pharm Res 30, 117–130 (2013). https://doi.org/10.1007/s11095-012-0854-x

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  • DOI: https://doi.org/10.1007/s11095-012-0854-x

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