Advertisement

Pharmaceutical Research

, 35:157 | Cite as

Rotational Rheology of Bovine Serum Albumin Solutions: Confounding Effects of Impurities, Mechanistic Considerations and Potential Implications on Protein Formulation Development

Research Paper

Abstract

Purpose

To show and rationalize the confounding effects on the rotational/oscillatory rheology of surface active impurities in commercial protein formulations such as bovine serum albumin, BSA.

Methods

Bulk and interfacial rotational/oscillatory rheology were used to study the viscosity, complex viscosity, storage/elastic modulus, G’ and loss/viscous modulus, G”, as a function of time of aqueous formulations of BSA and their purified components.

Results

Viscosity/time profiles at steady shear for different commercial BSA products and lots showed viscosity increase, decrease and time-independent profiles at low shear rates. All lots showed shear thinning. BSA monomer and dimers/aggregates, in general, showed similar profiles. Addition of low levels of surfactant or high shear rates rendered all solutions to be Newtonian-like. Interfacial viscosity studies paralleled those on the rotational rheometer. G’ > G” with viscosity increase and G’ < G” with viscosity decrease over time.

Conclusions

We provide a rational explanation for the time-dependent and source-dependent rheological behavior of aqueous formulations of commercially available BSA proteins based on the migration of protein and surface active impurities to the air/water interface within the rheometer plates leading to the formation and breakdown of protein networks. Highly purified proteins is warranted in rheological studies of protein drug product candidates.

KEY WORDS

bovine serum albumin high molecular weight species/aggregates mechanism protein stability rotational/interfacial rheology surface active impurity 

Abbreviations

BSA

Bovine serum albumin

DWR

Double-wall ring

HMWS

High molecular weight species

MS

Mass spectrometry

mVROC

microfluidic viscometer-rheometer-on-chip

MW

Molecular weight

Polysorbate 20

PS20

RP-HPLC

Reversed phase high-performance Liquid chromatography

SE-HPLC

Size exclusion high-performance Liquid chromatography

SANS

Small angle Neutron scattering

Notes

ACKNOWLEDGMENTS AND DISCLOSURES

The authors wish to thank Alice Beekman for her pioneering work on the rheology of protein and polymer formulations and who brought to our attention the possibility of proteins at surfaces contributing to the overall rheology she observed; in addition we thank her for her contributions to the manuscript. The authors are grateful to Dave Brems and Margaret Ricci for support of this research; to Da Ren for technical support in LC/MS analysis; to Lyanne Wong for her early work on protein rheology. The authors also thank Philip Rolfe and David Bohnsack for their contributions to this work.

Supplementary material

11095_2018_2423_MOESM1_ESM.doc (3.9 mb)
ESM 1 (DOC 3982 KB)

References

  1. 1.
    Shire SJ, Shahrokh Z, Liu J. Biotechnol.: Pharm. Aspects. 2010;11:131–47.Google Scholar
  2. 2.
    Harris RJ, Shire SJ, Winter C. Commercial manufacturing scale formulation and analytical characterization of therapeutic recombinant antibodies. Drug Dev Res. 2004;61:137–54.CrossRefGoogle Scholar
  3. 3.
    Jezek J, Rides M, Derham B, Moore J, Cerasoli E, Simler R, et al. Viscosity of concentrated therapeutic protein compositions. Adv Drug Deliv Rev. 2011;63:1107–17.CrossRefPubMedGoogle Scholar
  4. 4.
    Galush WJ, Le LN, Moore JMR. Viscosity Behavior of High-Concentration Protein Mixtures. J Pharm Sci. 2012;101:1012–20.CrossRefPubMedGoogle Scholar
  5. 5.
    Yadav S, Shire SJ, Kalonia DS. Viscosity Analysis of High Concentration Bovine Serum Albumin Aqueous Solutions. Pharm Res. 2011;28:1973–83.CrossRefPubMedGoogle Scholar
  6. 6.
    Oates KMN, Krause WE, Jones RL, Colby RH. Rheopexy of synovial fluid and protein aggregation. J R Soc Interface. 2006;3:167–74.CrossRefPubMedGoogle Scholar
  7. 7.
    Cascao Pereira LG, Theodoly O, Blanch HW, Radke CJ. Dilatational Rheology of BSA Conformers at the Air/Water Interface. Langmuir. 2003;19:2349–56.CrossRefGoogle Scholar
  8. 8.
    Grigoriev DO, Derkatch S, Kraegel J, Miller R. Relationship between structure and rheological properties of mixed BSA/Tween 80 adsorption layers at the air/water interface. Food Hydrocoll. 2007;21:823–30.CrossRefGoogle Scholar
  9. 9.
    Sharma V, Jaishankar A, Wang Y-C, McKinley GH. Rheology of globular proteins: apparent yield stress, high shear rate viscosity and interfacial viscoelasticity of bovine serum albumin solutions. Soft Matter. 2011;7:5150–60.CrossRefGoogle Scholar
  10. 10.
    Brownsey GJ, Noel TR, Parker R, Ring SG. The Glass Transition Behavior of the Globular Protein Bovine Serum Albumin. Biophys J. 2003;85:3943–50.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Ikeda S, Nishinari K. Intermolecular Forces in Bovine Serum Albumin Solutions Exhibiting Solidlike Mechanical Behaviors. Biomacromolecules. 2000;1:757–63.CrossRefPubMedGoogle Scholar
  12. 12.
  13. 13.
    Cohn EJ, Strong LE, Hughes WL Jr, Mulford DJ, Ashworth JN, Melin M, et al. Preparation and Properties of Serum and Plasma Proteins. IV. A System for the Separation into Fractions of the Protein and Lipoprotein Components of Biological Tissues and Fluids1a,b,c,d. J Am Chem Soc. 1946;68:459–75.CrossRefPubMedGoogle Scholar
  14. 14.
    Beaufils S, Hadaoui-Hammoutene R, Vie V, Miranda G, Perez J, Terriac E, et al. Comparative behaviour of goat β and αs1-caseins at the air–water interface and in solution. Food Hydrocoll. 2007;21:1330–43.CrossRefGoogle Scholar
  15. 15.
  16. 16.
    Vandebril S, Franck A, Fuller GG, Moldenaers P, Vermant J. A double wall-ring geometry for interfacial shear rheometry. Rheol Acta. 2010;49:131–44.CrossRefGoogle Scholar
  17. 17.
    Jaishankar A, Sharma V, McKinley GH. Interfacial viscoelasticity, yielding and creep ringing of globular protein–surfactant mixtures. Soft Matter. 2011;7:7623–34.CrossRefGoogle Scholar
  18. 18.
    Lawrence ASC, Needham J, Shen S-C. Studies on the anomalous viscosity and flow-birefringence of protein solutions: i. general behaviour of proteins subjected to shear. J Gen Physiol. 1944;27:201–32.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Castellanos MM, Pathak JA, Colby RH. Both protein adsorption and aggregation contribute to shear yielding and viscosity increase in protein solutions. Soft Matter. 2014;10:122–31.CrossRefPubMedGoogle Scholar
  20. 20.
    Serrien G, Geeraerts G, Ghosh L, Joos P. Dynamic surface properties of adsorbed protein solutions: BSA, casein and buttermilk. Colloids Surf. 1992;68:219–33.CrossRefGoogle Scholar
  21. 21.
    Murray BS. Rheological properties of protein films. Curr Opin Colloid Interface Sci. 2011;16:27–35.CrossRefGoogle Scholar
  22. 22.
    Semwogerere D, Weeks ER. Phys. Fluids. 2008;20:043306/043301–7.Google Scholar
  23. 23.
    Nossal R, Glinka CM, Chen SH. SANS studies of concentrated protein solutions. I. Bovine serum albumin. Biopolymers. 1986;25:1157–75.CrossRefPubMedGoogle Scholar
  24. 24.
    Porcar L, Falus P, Chen W-R, Faraone A, Fratini E, Hong K, et al. Formation of the Dynamic Clusters in Concentrated Lysozyme Protein Solutions. J Phys Chem Lett. 2010;1:126–9.CrossRefGoogle Scholar
  25. 25.
    Yearley EJ, Godfrin PD, Perevozchikova T, Zhang H, Falus P, Porcar L, et al. Observation of small cluster formation in concentrated monoclonal antibody solutions and its implications to solution viscosity. Biophys J. 2014;106:1763–70.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Valstar A, Almgren M, Brown W, Vasilescu M. The Interaction of Bovine Serum Albumin with Surfactants Studied by Light Scattering. Langmuir. 2000;16:922–7.CrossRefGoogle Scholar
  27. 27.
    Zadymova NM, Yampol'skaya GP, Filatova LY. Colloid J. 2006;68:162–72.CrossRefGoogle Scholar
  28. 28.
    Krishnan A, Sturgeon J, Siedlecki CA, Vogler EA. J. Biomed. Mater. Res., Part A. 2004;68A:544–57.CrossRefGoogle Scholar
  29. 29.
    Morris VJ, Gunning AP. Microscopy, microstructure and displacement of proteins from interfaces: implications for food quality and digestion. Soft Matter. 2008;4:943–51.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Drug Product TechnologyAmgen Inc.Thousand OaksUSA
  2. 2.Biological Relevance and CharacterizationAmgen Inc.Thousand OaksUSA
  3. 3.GoldenBiotech, LLCNewbury ParkUSA

Personalised recommendations