Development of a high-throughput solubility screening assay for use in antibody discovery

MAbs. 2019 May/Jun;11(4):747-756. doi: 10.1080/19420862.2019.1589851. Epub 2019 Mar 26.

Abstract

Poor solubility is a common challenge encountered during the development of high concentration monoclonal antibody (mAb) formulations, but there are currently no methods that can provide predictive information on high-concentration behavior of mAbs in early discovery. We explored the utility of methodologies used for determining extrapolated solubility as a way to rank-order mAbs based on their relative solubility properties. We devised two approaches to accomplish this: 1) vapor diffusion technique utilized in traditional protein crystallization practice, and 2) polyethylene glycol (PEG)-induced precipitation and quantitation by turbidity. Using a variety of in-house mAbs with known high-concentration behavior, we demonstrated that both approaches exhibited reliable predictability of the relative solubility properties of these mAbs. Optimizing the latter approach, we developed a format that is capable of screening a large panel of mAbs in multiple pH and buffer conditions. This simple, material-saving, high-throughput approach enables the selection of superior molecules and optimal formulation conditions much earlier in the antibody discovery process, prior to time-consuming and material intensive high-concentration studies.

Keywords: Solubility screening; formulation; high-throughput; mAb developability assessment.

MeSH terms

  • Antibodies, Monoclonal / chemistry*
  • Crystallization
  • HEK293 Cells
  • High-Throughput Screening Assays / methods*
  • Humans
  • Hydrogen-Ion Concentration
  • Polyethylene Glycols / chemistry
  • Solubility

Substances

  • Antibodies, Monoclonal
  • Polyethylene Glycols