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Protein A chromatography increases monoclonal antibody aggregation rate during subsequent low pH virus inactivation hold.

Mazzer AR, Perraud X, Halley J, O'Hara J, Bracewell DG - J Chromatogr A (2015)

Bottom Line: Yet, a more limited set of evidence suggests that low pH may not be the sole cause of aggregation in protein A chromatography, rather, other facets of the process may contribute significantly.Similar experiments were implemented in the absence of a chromatography step, i.e. IgG4 aggregation at low pH.Rate constants for aggregation after protein A chromatography were considerably higher than those from low pH exposure alone; a distinct shift in aggregation rates was apparent across the pH range tested.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemical Engineering, University College London, Bernard Katz Building, Gordon Street, London WC1H 0AH, United Kingdom.

No MeSH data available.


Related in: MedlinePlus

Y0 (plateau point parameter) from exponential decay curve fits plotted against pH at 0.9 mg/mL, 2.7 mg/mL and 4.5 mg/mL in solution, and at approximately 2.7 mg/mL after protein A chromatography. Y-error bars show the standard error for the Y0 parameter (Eq. (2)) obtained from least squares fitting; x-error bars represent pH measurement error. The data was pooled and fitted with a linear trend (solid line); error weighting was not used for the fitting. The 95% confidence interval is shown as dotted lines. The fit was significant with an adjusted r2 of 0.75 and P < 0.01.
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fig0030: Y0 (plateau point parameter) from exponential decay curve fits plotted against pH at 0.9 mg/mL, 2.7 mg/mL and 4.5 mg/mL in solution, and at approximately 2.7 mg/mL after protein A chromatography. Y-error bars show the standard error for the Y0 parameter (Eq. (2)) obtained from least squares fitting; x-error bars represent pH measurement error. The data was pooled and fitted with a linear trend (solid line); error weighting was not used for the fitting. The 95% confidence interval is shown as dotted lines. The fit was significant with an adjusted r2 of 0.75 and P < 0.01.

Mentions: In summary, the plateau point was not considered a robust parameter to quantify the aggregation that occurred under the given conditions. This is further illustrated in Fig. 6, where the relationship between pH, y0 values and IgG concentration for solution-only and column experiments is displayed. The general trend of decreasing monomer loss with increasing pH can be seen, and it is also apparent that the effects of IgG concentration and the column step on y0 are not well defined.


Protein A chromatography increases monoclonal antibody aggregation rate during subsequent low pH virus inactivation hold.

Mazzer AR, Perraud X, Halley J, O'Hara J, Bracewell DG - J Chromatogr A (2015)

Y0 (plateau point parameter) from exponential decay curve fits plotted against pH at 0.9 mg/mL, 2.7 mg/mL and 4.5 mg/mL in solution, and at approximately 2.7 mg/mL after protein A chromatography. Y-error bars show the standard error for the Y0 parameter (Eq. (2)) obtained from least squares fitting; x-error bars represent pH measurement error. The data was pooled and fitted with a linear trend (solid line); error weighting was not used for the fitting. The 95% confidence interval is shown as dotted lines. The fit was significant with an adjusted r2 of 0.75 and P < 0.01.
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4582070&req=5

fig0030: Y0 (plateau point parameter) from exponential decay curve fits plotted against pH at 0.9 mg/mL, 2.7 mg/mL and 4.5 mg/mL in solution, and at approximately 2.7 mg/mL after protein A chromatography. Y-error bars show the standard error for the Y0 parameter (Eq. (2)) obtained from least squares fitting; x-error bars represent pH measurement error. The data was pooled and fitted with a linear trend (solid line); error weighting was not used for the fitting. The 95% confidence interval is shown as dotted lines. The fit was significant with an adjusted r2 of 0.75 and P < 0.01.
Mentions: In summary, the plateau point was not considered a robust parameter to quantify the aggregation that occurred under the given conditions. This is further illustrated in Fig. 6, where the relationship between pH, y0 values and IgG concentration for solution-only and column experiments is displayed. The general trend of decreasing monomer loss with increasing pH can be seen, and it is also apparent that the effects of IgG concentration and the column step on y0 are not well defined.

Bottom Line: Yet, a more limited set of evidence suggests that low pH may not be the sole cause of aggregation in protein A chromatography, rather, other facets of the process may contribute significantly.Similar experiments were implemented in the absence of a chromatography step, i.e. IgG4 aggregation at low pH.Rate constants for aggregation after protein A chromatography were considerably higher than those from low pH exposure alone; a distinct shift in aggregation rates was apparent across the pH range tested.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemical Engineering, University College London, Bernard Katz Building, Gordon Street, London WC1H 0AH, United Kingdom.

No MeSH data available.


Related in: MedlinePlus