Antibody aggregates
Posted: Fri Oct 16, 2009 3:40 pm
Anyone aware of an article that compares direct injections with diluted injections for characterizing of antibodies aggregates? thanks
The first one is the most applicable and the others will basically show you that there is a lot more to learn!
Basically if you have doubt about the separation process either not seeing or the dilution breaking up the aggregates then you need to investigate both mass recovery on the column (ie how much did you inject and how much did you get out!) and also differences in initial concentration, injection volume and also flow rate as this affects the shear force...........
what I am really saying is that this is a lot of work and you may also need some additional detectors - Multi-angle light scattering and Analytical Ultra Centrifuge to do the job properly.
J. S. Bee, J. L. Stevenson, B. Mehta, J. Svitel, J. Pollastrini, R. Platz, E. Freund, J. F. Carpenter, T. W. Randolph, "Response of a Concentrated Monoclonal Antibody Formulation to High Shear", Biotechnology and Bioengineering 103(5), 936-943 (2009).
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A. Hawe, W. Friess, M. Sutter, W. Jiskoot, "Online fluorescent dye detection method for the characterization of immunoglobulin G aggregation by size exclusion chromatography and asymmetrical flow field flow fractionation", Analytical Biochemistry 378(2), 115-122 (2008).
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B. Demeule, M. J. Lawrence, A. F. Drake, R. Gurny, T. Arvinte, "Characterization of protein aggregation: the case of a therapeutic immunoglobulin", Biochimica et Biophysica Acta 1774(1), 146-153 (2007).
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J. P. Gabrielson, M. L. Brader, A. H. Pekar, K. B. Mathis, G. Winter, J. F. Carpenter, T. W. Randolph, "Quantitation of Aggregate Levels in a Recombinant Humanized Monoclonal Antibody Formulation by Size-Exclusion Chromatography, Asymmetrical Flow Field-Flow Fractionation, and Sedimentation Velocity", Journal of Pharmaceutical Sciences 96(2), 268-279 (2007).