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An alternative assay to hydrophobic interaction chromatography for high-throughput characterization of monoclonal antibodies
- Source :
- mAbs
- Publication Year :
- 2015
- Publisher :
- Taylor & Francis, 2015.
-
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 concentration. We find that hydrophobic mAbs, as identified by HIC, generally show significant self-association at low to moderate ammonium sulfate concentrations, while hydrophilic mAbs typically show self-association only at high ammonium sulfate concentrations. The correlation between AC-SINS and HIC measurements suggests that our assay, which can evaluate tens to hundreds of mAbs in a parallel manner and requires only small (microgram) amounts of antibody, will enable early identification of mAb candidates with low hydrophobicity and improved biophysical properties.
- Subjects :
- Ammonium sulfate
medicine.drug_class
High-throughput screening
Immunology
Metal Nanoparticles
Monoclonal antibody
high-throughput screening
Hydrophobic effect
chemistry.chemical_compound
developability
medicine
Immunology and Allergy
Humans
Solubility
HIC
Chromatography
biology
Hydrophilic interaction chromatography
solubility
Spectrum Analysis
aggregation
Antibodies, Monoclonal
hydrophobic interaction chromatography
HEK293 Cells
chemistry
Polyclonal antibodies
Colloidal gold
viscosity
biology.protein
manufacturability
Gold
Hydrophobic and Hydrophilic Interactions
Reports
self-interaction nanoparticle spectroscopy
self-association
Chromatography, Liquid
Subjects
Details
- Language :
- English
- ISSN :
- 19420870 and 19420862
- Volume :
- 7
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- mAbs
- Accession number :
- edsair.doi.dedup.....b67c86a9d1cb8beec138b48893a3f554