Mid-infrared spectroscopy-based antibody aggregate quantification in cell culture fluids.

Therapeutic antibody purification involves several steps which potentially induce antibody aggregation. Currently, aggregate monitoring mainly employs chromatographic, SDS-PAGE and light scattering techniques. In this study, the feasibility of mid-infrared spectroscopy (MIR) for the quantification of soluble antibody aggregates was investigated. Several multivariate models were evaluated to quantify antibody aggregation in chromatography elution streams and in clarified CHO cell culture supernatants (a surrogate for bioreactor output). A general model was established that is applicable for aggregate quantification directly from different cell culture solutions. Real-process samples and process-sample mimics were used to verify the general aggregate quantification model using two different antibodies. Results showed good prediction ability down to 1% aggregate content. Together with recently published results using MIR for host cell protein and target protein quantification, the results presented here indicate that MIR could provide multi-parameter process information from a single, fast, cost-effective and straightforward measurement. In conclusion, our study demonstrates that MIR is suitable for aggregate quantification in therapeutic antibody purification processes.