Dept Biochemical Engineering, UCL, London UK – p.dalby@ucl.ac.uk

Screen early – fail early: Rapid analytical screens for protein formulation

Biophysical analyses are essential for understanding the effect of formulation excipients on protein stability. However, most are not accessible for small quantities of precious materials or for higher-throughput analyses. Microscale and microfluidic platforms are enabling small quantities of proteins to be rapidly analysed under pre-formulation, stress and bioprocess conditions, to assess their stability. Automatable high-throughput methods for measuring protein thermostability, tolerance to freeze-drying, solubility, aggregation and precipitation will be introduced. A recently established rapid and accurate microfluidics-based biophysical analysis of protein stability and ligand interactions will also be discussed.

{aridoc engine=”google” width=”600″ height=”700″}images/mibio2012/speakers/paul-dalby.pdf{/aridoc}

[1] Aucamp, J.P., Martinez-Torres, R.J., Hibbert, E.G. and Dalby, P.A. (2008) Biotech. Bioeng. 99, 1303-1310. A microplate-based evaluation of complex denaturation pathways: Structural stability of E. coli transketolase

[2] Aucamp, J. P., Cosme, A. M., Lye, G. J. and Dalby, P. A. (2005) Biotech Bioeng. 89 (5), 599-607. High-throughput measurement of protein stability in microtiter plates

[3] Grant, Y., Matejtschuk, P., Dalby, P.A. (2009) Biotech. Bioeng. 104(5), 957-964. Rapid optimisation of protein freeze-drying formulations using ultra scale-down and factorial design of experiment in microplates.

[4] Ahmad, S.S., Dalby, P.A. (2010) Biotechnol. Bioeng. 108(2):322-332. Thermodynamic parameters for salt-induced reversible protein precipitation from automated microscale experiments.

[5] Gaudet, M., Remtulla, N., Jackson, S.E., Main, E.R.G., Bracewell, D.G., Aeppli, G., Dalby, P.A. (2010) Protein Science. 19, 1544-1554. Protein denaturation and protein:drug interactions from intrinsic protein fluorescence measurements at the nanolitre scale.

[6] Ordidge G.C., Mannall, G., Liddell, J., Dalby, P.A., Micheletti, M. (2012) Biotechnology Progress 28(2):435-444. A generic hierarchical screening method for the analysis of microscale refolds using an automated robotic platform.

[7] Grant, Y., Matejtschuk, P., Bird, C., Wadhwa, M., Dalby, P.A. (2012) Biotechnol. Letts. 34(4):641-648. Rapid identification and optimisation of freeze drying formulation using microscale and design of experiment approaches: A case study using granulocyte colony stimulating factor (G-CSF).

[8] Abe, Y., Gor, J., Bracewell, D.G., Perkins, S.J., Dalby, P.A. (2010) Biochem. J. 432(1):101-111. Masking of the Fc region in human IgG4 by constrained X-ray scattering modelling: implications for antibody function and therapy.