A CFD model for predicting protein aggregation in low-pH virial
inactivation for mAb production
Abstract
Significant amounts of soluble product aggregates were observed in the
low-pH viral inactivation (VI) opertion during an initial scale-up run
for an IgG4 monoclonal antibody (mAb IgG4-N1). Being earlier in
development, a scale-down model did not exist, nor was it practical to
use costly Protein A eluate (PAE) for testing the VI process at scale,
thus, a computational fluid dynamics (CFD)-based high molecular weight
(HMW) prediction model was developed for troubleshooting and risk
mitigation. It was previously reported that the IgG4-N1 molecules upon
exposure to low pH tend to change into transient and partially unfolded
monomers during VI acidification (i.e., VIA) and form aggregates after
neutralization (i.e., VIN) (Jin et al. 2019). Therefore, the CFD model
reported here focuses on the VIA step. The model mimics the continuous
addition of acid to PAE and tracks acid distribution during VIA. Based
on the simulated low-pH zone (≤ pH 3.3) profiles and PAE properties, the
integrated low-pH zone (ILPZ) value was obtained to predict HMW level at
the VI step. The simulations were performed to examine the operating
parameters, such as agitation speed, acid addition rate, and protein
concentration of PAE, of the pilot scale (50-200L) runs. The conditions
with predictions of no product aggregation risk were recommended to the
real scale-up runs, resulted in 100% success rate of the consecutive 12
pilot-scale runs. This work demonstrated that the CFD-based HMW
prediction model could be used as a tool to facilitate the scale up of
the low-pH VI process directly from bench to pilot/production scale.