An alternative assay to hydrophobic interaction chromatography for high-throughput characterization of monoclonal antibodies

MAbs
Patricia EstepYingda Xu

Abstract

The effectiveness of therapeutic monoclonal antibodies (mAbs) is governed not only by their bioactivity, but also by their biophysical properties. Assays for rapidly evaluating the biophysical properties of mAbs are valuable for identifying those most likely to exhibit superior properties such as high solubility, low viscosity and slow serum clearance. Analytical hydrophobic interaction chromatography (HIC), which is performed at high salt concentrations to enhance hydrophobic interactions, is an attractive assay for identifying mAbs with low hydrophobicity. However, this assay is low throughput and thus not amenable to processing the large numbers of mAbs that are commonly generated during antibody discovery. Therefore, we investigated whether an alternative, higher throughput, assay could be developed that is based on evaluating antibody self-association at high salt concentrations using affinity-capture self-interaction nanoparticle spectroscopy (AC-SINS). Our approach is to coat gold nanoparticles with polyclonal anti-human antibodies, use these conjugates to immobilize human mAbs, and evaluate mAb self-interactions by measuring the plasmon wavelengths of the antibody conjugates as a function of ammonium sulfate concentrati...Continue Reading

References

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Citations

Jan 18, 2017·Proceedings of the National Academy of Sciences of the United States of America·Tushar JainK Dane Wittrup
Dec 12, 2018·Biotechnology and Bioengineering·Feifan DuBenjamin J Hackel
Feb 17, 2019·Protein Engineering, Design & Selection : PEDS·Lilia A RabiaPeter M Tessier
Jul 1, 2020·BioMed Research International·Anthony Mackitz DzisooJian Huang
Oct 16, 2020·International Journal of Molecular Sciences·Manali S SawantPeter M Tessier
Jun 4, 2021·Proceedings of the National Academy of Sciences of the United States of America·Alexander W GolinskiBenjamin J Hackel
Jun 29, 2021·Journal of Pharmaceutical Sciences·Yuhong ZengDavid S Soane
Oct 16, 2019·Molecular Pharmaceutics·Priyanka SinghRobin Curtis
Sep 14, 2018·Analytical Chemistry·Li ZhouWilliam Blaine Stine

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Methods Mentioned

BETA
cross-interaction chromatography
ELISA
fluorescence-activated cell sorting
affinity-capture
light scattering

Software Mentioned

excel
SINS
AC

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