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High-Throughput Analytical Light Scattering for Protein Quality Control and Characterization

  • Daniel SomeEmail author
  • Vladimir Razinkov
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2025)

Abstract

We present a review of high-throughput techniques for the characterization and quality control of proteins in the course of purification, evaluation, and formulation, based on static and dynamic light scattering. Multi-angle static light scattering (MALS) in combination with rapid, low-volume UHPLC size exclusion chromatography is effective in characterizing key biophysical properties, while dynamic light scattering (DLS) in high-throughput microwell-plate format provides large quantities of data in a short time to screen many conditions, excipients, cell lines, or candidate biotherapeutics.

Key words

SEC-MALS Dynamic light scattering Protein quality Aggregation Protein formulation Monoclonal antibodies 

References

  1. 1.
    Liu T, Chu B (2015) Light-scattering by proteins. In: Somasundaran P (ed) Encyclopedia of surface and colloid science. CRC, Boca RatonGoogle Scholar
  2. 2.
    Folta-Stogniew EJ (2009) Macromolecular interactions: light scattering. In: Encyclopedia of life sciences. Wiley, HobokenGoogle Scholar
  3. 3.
    PJ W (1993) Light scattering and the absolute characterization of macromolecules. Anal Chim Acta 272:1–40CrossRefGoogle Scholar
  4. 4.
    Zhao H, Brown PH, Schuck P (2011) On the distribution of protein refractive index increments. Biophys J 100(9):2309–2317CrossRefGoogle Scholar
  5. 5.
    Some D, Kenrick S (2012) Characterization of protein-protein interactions via static and dynamic light scattering. In: Cai J (ed) Protein interactions. In Tech, RijekaGoogle Scholar
  6. 6.
    Wen J, Arakawa T, Philo JS (1996) Size-exclusion chromatography with on-line light-scattering, absorbance, and refractive index detectors for studying proteins and their interactions. Anal Biochem 240(2):155–166CrossRefGoogle Scholar
  7. 7.
    Gimpl K, Klement J, Keller S (2016) Characterising protein/detergent complexes by triple-detection size-exclusion chromatography. Biol Proced Online 18(1):4CrossRefGoogle Scholar
  8. 8.
    Rebolj K, Pahovnik D, Žagar E (2012) Characterization of a protein conjugate using an asymmetrical-flow field-flow fractionation and a size-exclusion chromatography with multi-detection system. Anal Chem 84(17):7374–7383CrossRefGoogle Scholar
  9. 9.
    Bouvier ESP, Koza SM (2014) Advances in size-exclusion separations of proteins and polymers by UHPLC. TrAC Trends Anal Chem 63:85–94CrossRefGoogle Scholar
  10. 10.
    Philo JS (2009) A critical review of methods for size characterization of non-particulate protein aggregates. Curr Pharm Biotechnol 10(4):359–372CrossRefGoogle Scholar
  11. 11.
    He F et al (2010) High-throughput dynamic light scattering method for measuring viscosity of concentrated protein solutions. Anal Biochem 399(1):141–143CrossRefGoogle Scholar
  12. 12.
    Menzen T, Friess W (2014) Temperature-ramped studies on the aggregation, unfolding, and interaction of a therapeutic monoclonal antibody. J Pharm Sci 103(2):445–455CrossRefGoogle Scholar
  13. 13.
    Lehermayr C et al (2011) Assessment of net charge and protein–protein interactions of different monoclonal antibodies. J Pharm Sci 100(7):2551–2562CrossRefGoogle Scholar
  14. 14.
    Kenrick S, Some D (2018) The diffusion interaction parameter kd as an indicator of colloidal and thermal stability. Available from http://www.wyatt.com/files/literature/app-notes/dls-plate/WP5004-diffusion-interaction-parameter-for-colloidal-and-thermal-stability.pdf
  15. 15.
    He F et al (2011) High-throughput assessment of thermal and colloidal stability parameters for monoclonal antibody formulations. J Pharm Sci 100(12):5126–5141CrossRefGoogle Scholar
  16. 16.
    Austerberry JI et al (2017) The effect of charge mutations on the stability and aggregation of a human single chain Fv fragment. Eur J Pharm Biopharm 115:18–30CrossRefGoogle Scholar
  17. 17.
    Connolly BD et al (2012) Weak interactions govern the viscosity of concentrated antibody solutions: high-throughput analysis using the diffusion interaction parameter. Biophys J 103(1):69–78CrossRefGoogle Scholar
  18. 18.
    Dear BJ et al (2017) Contrasting the influence of cationic amino acids on the viscosity and stability of a highly concentrated monoclonal antibody. Pharm Res 34(1):193–207CrossRefGoogle Scholar
  19. 19.
    He F et al (2011) Screening of monoclonal antibody formulations based on high-throughput thermostability and viscosity measurements: design of experiment and statistical analysis. J Pharm Sci 100(4):1330–1340CrossRefGoogle Scholar
  20. 20.
    Luo H et al (2017) Liquid-liquid phase separation causes high turbidity and pressure during low pH elution process in Protein A chromatography. J Chromatogr A 1488:57–67CrossRefGoogle Scholar
  21. 21.
    Some D (2014) Protein quality control in SPR and BLI high-throughput screening studies. Available from http://www.wyatt.com/files/literature/white-papers/protein-quality-control-high-throughput-screening-studies.pdf

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Wyatt Technology Corp.Santa BarbaraUSA
  2. 2.Drug Product Technologies, Amgen, Inc.Thousand OaksUSA

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