458834 Pre-Formulation Tool to Rapidly Predict the Stability of Monoclonal Antibodies

Monday, November 14, 2016
Grand Ballroom B (Hilton San Francisco Union Square)
Sarah Hedberg1, Daryl Williams1, Jerry Heng1 and John Liddell2, (1)Department of Chemical Engineering, Imperial College London, London, United Kingdom, (2)JML Biotechnology Consulting, Cleveland, United Kingdom

Pre-formulation tool to rapidly predict the stability of monoclonal antibodies

S. Hedberg*, J.Y.Y Heng*, D.R. Williams* and J. Liddell**

ABSTRACT

Protein-protein molecular interactions are known to be involved in protein solution aggregation behaviour and are a common issue for the manufacturing of therapeutic proteins such as mAbs. Much effort has been employed to gain a better understanding of aggregation. However, the mechanisms leading to protein aggregation are still not fully understood. The osmotic second virial coefficient (B22) is a fundamental physiochemical property that describes protein-protein interactions solution, which can be a useful tool for predicting the aggregation propensity of proteins.

One way of predicting aggregation propensity is self-interaction chromatography (SIC), which has demonstrated recent promise as a tool for the better understanding of phase behaviour of proteins. Another technique, cross-interaction chromatography (CIC), has shown to be an even more high-throughput technique than its predecessor with possibly the same capabilities.

The first part of this paper is a scale-down SIC study of therapeutic mAbs from laboratory scale macro-columns to micro-scale columns, which enables the use of micrograms or milligrams of mAb for B22 determinations in order to obtain a complete formulation screen.

The second part of this work presents an extensive formulation study of a mAbs system, varying pH and salt, as well as the presence of different stabilisers and different external factors known to induce aggregation. The B22 and B23 values determined from the formulation study are then correlated with aggregation kinetics obtained from size-exclusion chromatography and dynamic light scattering. It was shown that over all test conditions good correlations could especially be found between B22 and aggregation rate.

The third part of the study includes investigation of conformational changes of the monoclonal antibodies. The B22 data previously obtained were directly correlated with the conformational changes observed in the mAb to create a predictive tool that can detect any early conformational changes.

In summary this work intends to gain a better understanding of the fundamental mechanisms of aggregation and to develop a more predictive capability by studying the chromatogram output resulting from the self- and cross-interactions between the same or different proteins.

REFERENCES

Hedberg, S. H. M., Heng, J. Y. Y., Williams, D. R. & Liddell, J. M. (2016). Micro Scale Self-Interaction Chromatography of Proteins: A Mab Case-Study. Journal of Chromatography A, 1434, 57-63.

Quigley, A. & Williams, D. R. (2015). The Second Virial Coefficient as a Predictor of Protein Aggregation Propensity: A Self-Interaction Chromatography Study. European Journal of Pharmaceutics and Biopharmaceutics, 96, 282-290.


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