Predicting Protein-Protein Interactions of Concentrated Antibody Solutions Using Dilute Solution Data and...

Protein-protein interactions for solutions of an IgG1 molecule were quantified using static light scattering (SLS) measurements from low to high protein concentrations (c₂). SLS was used to determine second osmotic virial coefficients (B₂₂) at low c₂, and excess Rayleigh profiles (R^ex/K vs. c₂) and zero-q structure factors (S_q=0) as a function of c₂ at higher c₂ for a series of conditions (pH, sucrose concentration, and total ionic strength [TIS]). Repulsive (attractive) interactions were observed at low TIS (high TIS) for pH 5 and 6.5, with increasing repulsions when 5% w/w sucrose was also present. Previously developed and refined coarse-grained antibody models were used to fit model parameters from B₂₂ versus TIS data. The resulting parameters from low-c₂ conditions were used as the sole input to multiprotein Monte Carlo simulations to predict high-c₂R^ex/K and S_q=0 behavior up to 150 g/L. Experimental results at high-c₂ conditions were quantitatively predicted by the simulations for the coarse-grained models that treated antibody molecules as either 6 or 12 (sub) domains, which preserved the basic shape of a monoclonal antibody. Finally, preferential accumulation of sucrose around the protein surface was identified via high-precision density measurements, which self-consistently explained the simulation and experimental SLS results.