Skip to main content
SpringerLink
Log in

Ionomer and protein size analysis by analytical ultracentrifugation and electrospray scanning mobility particle sizer

  • Original Article
  • Published:
European Biophysics Journal Aims and scope Submit manuscript

Abstract

By combining analytical ultracentrifugation (AUC) in liquid phase and scanning mobility particle sizer (SMPS) in the gas phase, additional information on the particle size and morphology has been obtained for rigid particles. In this paper, we transfer this concept to soft particles, allowing us to analyze the size and molar mass of the short side chain perfluorosulfonic acid ionomer Aquivion® in a dilute aqueous suspension. The determination of the primary size and exact molar mass of this class of polymers is challenging since they are optically transparent and due to the formation of different aggregate structures depending on the concentration and solvent properties. First, validation of AUC and SMPS measurements was carried out using the well-defined biopolymers bovine serum albumin (BSA) and lysozyme (LYZ) to confirm the reliability of the results of the two unique and independent classifying methods. Then, the ionomer Aquivion® was studied using both techniques. From the mean molar mass of 185 ± 14 kDa obtained by AUC, a mean hydrodynamic diameter of 7.6 ± 0.5 nm was calculated. The particle size obtained from SMPS (7.1 nm) agrees very well with the results from AUC showing that the molecule was transferred into the gas phase without significantly changing its structure. In conclusion, the Aquivion® is molecularly dispersed in the used aqueous buffer solution without any aggregate formation in the investigated concentration range (< 2 g l−1).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Aldebert P, Dreyfus B, Pineri M (1986) Small-angle neutron scattering of perfluorosulfonated ionomers in solution. Macromolecules 19:2651

    Article  CAS  Google Scholar 

  • Aldebert P, Dreyfus B, Gebel G et al (1988) Rod like micellar structures in perfluorinated ionomer solutions. J Phys France 49:2101

    Article  CAS  Google Scholar 

  • Bailey M, Angley L, Perugini M (2009) Methods for sample labeling and meniscus determination in the fluorescence-detected analytical ultracentrifuge. Anal Biochem 390:218

    Article  CAS  Google Scholar 

  • Bhattacharyya S, Maciejewska P, Börger L et al (2006) Development of a fast fiber based UV-vis multiwavelength detector for an ultracentrifuge. In: Wandrey C, Cölfen H (eds) Analytical ultracentrifugation, VIII edn. Springer, New York, p 9

    Google Scholar 

  • Brookes E, Cao W, Demeler B (2010) A two-dimensional spectrum analysis for sedimentation velocity experiments of mixtures with heterogeneity in molecular weight and shape. Eur Biophys J 39:405

    Article  Google Scholar 

  • Canfield R (1963) The amino acid sequence of egg white lysozyme. J Biol Chem 238:2698

    CAS  PubMed  Google Scholar 

  • Carney R, Kim J, Qian H et al (2011) Determination of nanoparticle size distribution together with density or molecular weight by 2D analytical ultracentrifugation. Nat Commun 2:335

    Article  Google Scholar 

  • Cauchy A-L (1832) Mémoire sur la rectification des courbes et la quadrature des surfaces courbes. Oxford University, Oxford

    Google Scholar 

  • Cölfen H, Pauck T (1997) Determination of particle size distributions with angström resolution. Colloid Polym Sci 275:175

    Article  Google Scholar 

  • Colvin J (1952) The size and shape of lysozyme. Can J Chem 30:831

    Article  CAS  Google Scholar 

  • Demeler B (2005) UltraScan: a comprehensive data analysis software package for analytical ultracentrifugation experiments. In: Scott DJ, Harding SE, Rowe AJ (eds) Analytical ultracentrifugation: techniques and methods. Royal Society of Chemistry, London, p 210

    Google Scholar 

  • Demeler B, van Holde K (2004) Sedimentation velocity analysis of highly heterogeneous systems. Anal Biochem 335:279

    Article  CAS  Google Scholar 

  • Demeule B, Shire S, Liu J (2009) A therapeutic antibody and its antigen form different complexes in serum than in phosphate-buffered saline: a study by analytical ultracentrifugation. Anal Biochem 388:279

    Article  CAS  Google Scholar 

  • Elzey S, Grassian V (2010) Agglomeration, isolation and dissolution of commercially manufactured silver nanoparticles in aqueous environments. J Nanopart Res 12:1945

    Article  CAS  Google Scholar 

  • Epstein P (1924) On the Resistance Experienced by Spheres in their Motion through Gases. Phys Rev 23:710

    Article  CAS  Google Scholar 

  • Fasman G (1976) Handbook of biochemistry and molecular biology: proteins, 3rd edn. CRC Press, Cleveland

    Google Scholar 

  • Gebel G (2000) Structural evolution of water swollen perfluorosulfonated ionomers from dry membrane to solution. Polymer 41:5829

    Article  CAS  Google Scholar 

  • Ghielmi A, Vaccarono P, Troglia C et al (2005) Proton exchange membranes based on the short-side-chain perfluorinated ionomer. J Power Sources 145:108

    Article  CAS  Google Scholar 

  • Grot W (1986) Nafion as a separator in electrolyte cells in Nafion product bulletin. DuPont Co., Wilmington

    Google Scholar 

  • Harding S, Schuck P, Abdelhameed A et al (2011) Extended Fujita approach to the molecular weight distribution of polysaccharides and other polymeric systems. Methods 54:136

    Article  CAS  Google Scholar 

  • Hirayama K, Akashi S, Furuya M et al (1990) Rapid confirmation and revision of the primary structure of bovine serum albumin by ESIMS and Frit-FAB LC/MS. Biochem Biophys Res Commun 173:639

    Article  CAS  Google Scholar 

  • Kaddis C, Lomeli S, Yin S et al (2007) Sizing large proteins and protein complexes by electrospray ionization mass spectrometry and ion mobility. J Am Soc Mass Spectrom 18:1206

    Article  CAS  Google Scholar 

  • Kaufman S, Kuchumov A, Kazakevich M et al (1998) Analysis of a 3.6-MDa hexagonal bilayer hemoglobin from Lumbricus terrestris using a gas-phase electrophoretic mobility molecular analyzer. Anal Biochem 259:195

    Article  CAS  Google Scholar 

  • Kemptner J, Marchetti-Deschmann M, Siekmann J et al (2010) GEMMA and MALDI-TOF MS of reactive PEGs for pharmaceutical applications. J Pharm Biomed Anal 52:432

    Article  CAS  Google Scholar 

  • Koestner R, Roiter Y, Kozhinova I et al (2011) AFM imaging of adsorbed Nafion polymer on mica and graphite at molecular level. Langmuir 27:10157

    Article  CAS  Google Scholar 

  • Kratky O, Leopold H, Stabinger H (1973) The determination of the partial specific volume of proteins by the mechanical oscillator technique in part D: enzyme structure. Elsevier 27:98

    CAS  Google Scholar 

  • Ku B, de la Mora JF (2009) Relation between electrical mobility, mass, and size for nanodrops 1–6.5 nm in diameter in air. Aerosol Sci Technol 43:241

    Article  CAS  Google Scholar 

  • Lebowitz J, Lewis S, Schuck P (2002) Modern analytical ultracentrifugation in protein science: a tutorial review. Protein Sci 11:2067

    Article  CAS  Google Scholar 

  • Li H, Schlick S (1995) Effect of solvents on phase separation in perfluorinated ionomers, from electron spin resonance of VO2+ in swollen membranes and solutions. Polymer 36:1141

    Article  CAS  Google Scholar 

  • Liu W-H, Yu T-Y, Yu T et al (2007) Static light scattering and transmission microscopy study of dilute Nafion solutions. e-Polymers 109:1

    Google Scholar 

  • Lousenberg R (2005) Molar mass distributions and viscosity behavior of perfluorinated sulfonic acid polyelectrolyte aqueous dispersions. J Polym Sci 43:421

    Article  CAS  Google Scholar 

  • Mächtle W, Börger L (2006) Analytical ultracentrifugation of polymers and nanoparticles. Springer, Berlin Heidelberg

    Google Scholar 

  • Mauritz K, Moore R (2004) State of understanding of Nafion. Chem Rev 104:4535

    Article  CAS  Google Scholar 

  • Moore R, Martin C (1988) Chemical and morphological properties of solution-cast perfluorosulfonate ionomers. Macromolecules 21:1334

    Article  CAS  Google Scholar 

  • Mourey T, Slater L, Galipo R et al (2011) Size-exclusion chromatography of perfluorosulfonated ionomers. J Chromatogr A 1218:5801

    Article  CAS  Google Scholar 

  • Ngo T, Yu T, Lin H-L (2013) Influence of the composition of isopropyl alcohol/water mixture solvents in catalyst ink solutions on proton exchange membrane fuel cell performance. J Power Sources 225:293

    Article  CAS  Google Scholar 

  • Pavlov G, Perevyazko I, Schubert U (2010) Velocity sedimentation and intrinsic viscosity analysis of polystyrene standards with a wide range of molar masses. Macromol Chem Phys 211:1298

    Article  CAS  Google Scholar 

  • Perrin F (1936) Mouvement Brownien d’un ellipsoide (II). Rotation libre et dépolarisation des fluorescences. Translation et diffusion de molécules ellipsoidales. J Phys Radium 7:1

    Article  CAS  Google Scholar 

  • Pitschke M, Fels A, Schmidt B et al (1995) Polymeric fluorescent dyes for labeling of proteins and nucleic acids. Colloid Polym Sci 273:740

    Article  CAS  Google Scholar 

  • Planken K, Cölfen H (2010) Analytical ultracentrifugation of colloids. Nanoscale 2:1849

    Article  CAS  Google Scholar 

  • Saucy D, Ude S, Lenggoro I et al (2004) Mass analysis of water-soluble polymers by mobility measurement of charge-reduced ions generated by electrosprays. Anal Chem 76:1045

    Article  CAS  Google Scholar 

  • Schuck P (2000) Size-distribution analysis of macromolecules by sedimentation velocity ultracentrifugation and lamm equation modeling. Biophys J 78:1606

    Article  CAS  Google Scholar 

  • Schuck P, Zhao H, Brautigam C (2015) Basic principles of analytical ultracentrifugation. CRC Press, Boca Raton

    Book  Google Scholar 

  • Shibayama M, Matsunaga T, Kusano T et al (2014) SANS studies on catalyst ink of fuel cell. J Appl Polym Sci. https://doi.org/10.1002/app.39842

    Article  Google Scholar 

  • Siracusano S, Baglio V, Moukheiber E et al (2015) Performance of a PEM water electrolyser combining an IrRu-oxide anode electrocatalyst and a short-side chain Aquivion membrane. Int J Hydrogen Energy 40:14430

    Article  CAS  Google Scholar 

  • Strauss H, Karabudak E, Bhattacharyya S et al (2008) Performance of a fast fiber based UV/Vis multiwavelength detector for the analytical ultracentrifuge. Colloid Polym Sci 286:121

    Article  CAS  Google Scholar 

  • Tammet H (1995) Size and mobility of nanometer particles, clusters and ions. J Aerosol Sci 26:459

    Article  CAS  Google Scholar 

  • Tenzer S, Docter D, Rosfa S et al (2011) Nanoparticle size is a critical physicochemical determinant of the human blood plasma corona: a comprehensive quantitative proteomic analysis. ACS Nano 5:7155

    Article  CAS  Google Scholar 

  • Thajudeen T, Walter J, Srikantharajah R et al (2017) Determination of the length and diameter of nanorods by a combination of analytical ultracentrifugation and scanning mobility particle sizer. Nanoscale Horizons 2:253–260

    Article  CAS  Google Scholar 

  • Uchiyama S, Arisaka F, Stafford WF, Laue T (eds) (2016) Analytical ultracentrifugation. Instrumentation software, and applications. Springer, Tokyo

    Google Scholar 

  • Uttinger M, Walter J, Thajudeen T et al (2017) Brownian dynamics simulations of analytical ultracentrifugation experiments exhibiting hydrodynamic and thermodynamic non-ideality. Nanoscale 9:17770

    Article  CAS  Google Scholar 

  • van Holde K, Weischet W (1978) Boundary analysis of sedimentation-velocity experiments with monodisperse and paucidisperse solutes. Biopolymers 17:1387

    Article  Google Scholar 

  • van Holde K, Johnson W, Ho P (2006) Principles of physical biochemistry, 2nd edn. Pearson/Prentice Hall, Upper Saddle River

    Google Scholar 

  • Voigt M, Klaumünzer M, Thiem H et al (2010) Detailed analysis of the growth kinetics of ZnO nanorods in methanol. J Phys Chem C 114:6243

    Article  CAS  Google Scholar 

  • Walter J, Peukert W (2016) Dynamic range multiwavelength particle characterization using analytical ultracentrifugation. Nanoscale 8:7484

    Article  CAS  Google Scholar 

  • Walter J, Löhr K, Karabudak E et al (2014) Multidimensional analysis of nanoparticles with highly disperse properties using multiwavelength analytical ultracentrifugation. ACS Nano 8:8871

    Article  CAS  Google Scholar 

  • Walter J, Sherwood P, Lin W et al (2015) Simultaneous analysis of hydrodynamic and optical properties using analytical ultracentrifugation equipped with multiwavelength detection. Anal Chem 87:3396

    Article  CAS  Google Scholar 

  • Walter J, Segets D, Peukert W (2016) Extension of the deep UV-capabilities in multiwavelength spectrometry in analytical ultracentrifugation: the role of oil deposits. Part Part Syst Charact 33:184

    Article  Google Scholar 

  • Wang W, Damm C, Walter J et al (2016) Photobleaching and stabilization of carbon nanodots produced by solvothermal synthesis. Phys Chem Chem Phys 18:466

    Article  CAS  Google Scholar 

  • Welch C, Labouriau A, Hjelm R et al (2012) Nafion in dilute solvent systems: dispersion or solution? ACS Macro Lett. 1:1403

    Article  CAS  Google Scholar 

  • Wu X, Scott K, Puthiyapura V (2012) Polymer electrolyte membrane water electrolyser with Aquivion short side chain perfluorosulfonic acid ionomer binder in catalyst layers. Int J Hydrog Energy 37:13243

    Article  CAS  Google Scholar 

  • Xu F, Zhang H, Ilavsky J et al (2010) Investigation of a catalyst ink dispersion using both ultra-small-angle X-ray scattering and cryogenic TEM. Langmuir 26:19199

    Article  CAS  Google Scholar 

  • Yamaguchi M, Matsunaga T, Amemiya K et al (2014) Dispersion of rod-like particles of nafion in salt-free water/1-propanol and water/ethanol solutions. J Phys Chem B 118:14922

    Article  CAS  Google Scholar 

  • Yeo R (1980) Dual cohesive energy densities of perfluorosulfonic acid (Nafion) membrane. Polymer 21:432

    Article  CAS  Google Scholar 

  • Zhang C, Thajudeen T, Larriba C et al (2012) Determination of the scalar friction factor for nonspherical particles and aggregates across the entire knudsen number range by direct simulation monte carlo (DSMC). Aerosol Sci Technol 46:1065

    Article  CAS  Google Scholar 

  • Zhang H, Li J, Tang H et al (2014) Hydrogen crossover through perfluorosulfonic acid membranes with variable side chains and its influence in fuel cell lifetime. Int J Hydrog Energy 39:15989

    Article  CAS  Google Scholar 

  • Zhao H, Ghirlando R, Alfonso C et al (2015) A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation. PLoS ONE 10:e0126420

    Article  Google Scholar 

Download references

Acknowledgements

The authors acknowledge HI ERN and Bavaria (Grant-no. DBF01253) as well as Greenerity GmbH for financial support of this work. The authors further acknowledge the funding of the Deutsche Forschungsgemeinschaft (DFG) through the Cluster of Excellence “Engineering of Advanced Materials” as well as DFG Project PE 427/28-2. TT acknowledges the fellowship from Alexander von Humboldt foundation. SEW acknowledges the travel grant from ARBRE-MOBIEU and COST for the 23. International Analytical Ultracentrifugation Workshop and Symposium.

Author information

Author notes

  1. Thaseem Thajudeen

    Present address: School of Mechanical Sciences, Indian Institute of Technology Goa, Ponda, 403401, India

Authors and Affiliations

  1. Institute of Particle Technology (LFG), Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Cauerstrasse 4, 91058, Erlangen, Germany

    Simon E. Wawra, Martin Thoma, Johannes Walter, Christian Lübbert, Thaseem Thajudeen, Cornelia Damm & Wolfgang Peukert

  2. Interdisciplinary Center for Functional Particle Systems (FPS), Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Haberstrasse 9a, 91058, Erlangen, Germany

    Johannes Walter, Christian Lübbert, Thaseem Thajudeen & Wolfgang Peukert

Authors
  1. Simon E. Wawra
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Martin Thoma
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Johannes Walter
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christian Lübbert
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Thaseem Thajudeen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Cornelia Damm
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Wolfgang Peukert
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Wolfgang Peukert.

Additional information

Special Issue: 23rd International AUC Workshop and Symposium.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 2136 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wawra, S.E., Thoma, M., Walter, J. et al. Ionomer and protein size analysis by analytical ultracentrifugation and electrospray scanning mobility particle sizer. Eur Biophys J 47, 777–787 (2018). https://doi.org/10.1007/s00249-018-1314-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00249-018-1314-2

Keywords

Search

Navigation