Mapping the mAb Aggregation Propensity Using Self-Interaction Chromatography as a Screening Tool

• Published in: Analytical chemistry (Washington) 2018-03, Vol.90 (6), p.3878-3885
• Main Authors: Hedberg, Sarah H.M, Lee, DongKyu, Mishra, Yash, Haigh, Jonathan M, Williams, Daryl R
• Format: Article
• Language: English
• Subjects: Agglomeration; Antibodies, Monoclonal - chemistry; Chemistry; Chromatography; Chromatography, Gel - methods; Dynamic Light Scattering - methods; Experiments; Immobilized Proteins - chemistry; Immunoglobulin G - chemistry; Molecular interactions; Molecules; Photon correlation spectroscopy; Protein Aggregates; Protein Stability; Proteins; Scattering; Screening; Size exclusion chromatography; Sodium chloride; Sodium Chloride - chemistry; Stability; Temperature; Temperature dependence; Thermodynamics; Time dependence; Virial coefficients
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.7b04605

The osmotic second virial coefficient (B 2), which describes protein–protein molecular interactions in solution, was determined using self-interaction chromatography (SIC) for an IgG1-type mAb across a wide range of solution conditions. These data were compared with its time dependent aggregation behavior, as determined using size-exclusion chromatography (SEC), and its temperature dependent aggregation behavior using dynamic light scattering (DLS) over a four-week period (SEC) or overnight (DLS). DLS and SEC gave consistent data on aggregation behavior, which correlated well with experimental B 2 trends across the wide pH (4–9) and NaCl concentration (0–1.0 M) ranges studied. The IgG aggregated at pH 4 for 0.5–1.0 M NaCl concentrations and for 0 M NaCl concentrations at pH 8. Best stability against aggregation was exhibited for the pH range from 5 to 8 at 0.8–1.0 M NaCl. SIC data were able to be classified within the one-day solution conditions for aggregation, which were not identified for 2–3 weeks in the accelerated SEC stability study. The ability of SIC to provide such data rapidly reflects the fundamentally thermodynamic nature of this parameter and of the aggregation process itself. Proteins with attractive protein–protein interactions and negative B 2 coefficients in the range −3 to −6 clearly exhibit aggregation behavior, while B 2 values in the range 0 to 2 showed good stability toward aggregation. SIC allows the rapid screening of solution conditions for which mAbs will exhibit stability to aggregation while requiring 90% less time and material compared with that required for a conventional SEC aggregation study.