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Understanding the relationship between biotherapeutic protein stability and solid-liquid interfacial shear in constant region mutants of IgG1 and IgG4.

Tavakoli-Keshe R, Phillips JJ, Turner R, Bracewell DG - J Pharm Sci (2013)

Bottom Line: Results suggest that the techniques are orthogonal, with thermal methods based on intramolecular interaction and shear device stability based on localized unfolding revealing less stable regions that drive aggregation.Molecular modeling shows the modifications' effects on the antibody structures and indicates a possible role for Fc conformation and Fab-Fc docking in determining suspended protein stability.The data introduce the PDC value as an orthogonal stability indicator, complementary to traditional thermal methods, allowing lead antibody selection based on a more full understanding of process stability.

View Article: PubMed Central - PubMed

Affiliation: The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London, WC1E 7JE, UK; MedImmune, Granta Park, Cambridge, CB21 6GH, UK.

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(a) Comparison of PDC with Tm1 for all candidates at a disk rate of 9000 rpm. y-Axis error bars are for standard error of PDC fit, and x-axis error bars are for internal error of DSC. (b) Comparison of monomer remaining at week 4 in accelerated stability results, determined by SE-HPLC and the PDC as determined with custom shear device method for all candidates at a disk rate of 9000 rpm. x-Axis error bars are for standard error of PDC fit.
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fig07: (a) Comparison of PDC with Tm1 for all candidates at a disk rate of 9000 rpm. y-Axis error bars are for standard error of PDC fit, and x-axis error bars are for internal error of DSC. (b) Comparison of monomer remaining at week 4 in accelerated stability results, determined by SE-HPLC and the PDC as determined with custom shear device method for all candidates at a disk rate of 9000 rpm. x-Axis error bars are for standard error of PDC fit.

Mentions: The correlation of the thermal stability measured by DSC with conformational stability measured by adsorption to solid surfaces in a high-shear environment was assessed. Different antibodies are shown (Fig. 7a) in terms of their first melting temperature, Tm1, which represents unfolding of the CH2 region. This comparator is predominantly used in industrial screening of antibody candidates to determine the most stable candidates for accelerated stability studies. Comparison of the two stability indicators showed that there was no correlation between the thermal and the surface adsorption-based stability. The IgG4 candidates did show that although they were the least stable in the shear device, they had relatively good thermal stability for this transition. The lack of correlation suggests that different mechanisms of degradation are involved in the two testing methodologies.


Understanding the relationship between biotherapeutic protein stability and solid-liquid interfacial shear in constant region mutants of IgG1 and IgG4.

Tavakoli-Keshe R, Phillips JJ, Turner R, Bracewell DG - J Pharm Sci (2013)

(a) Comparison of PDC with Tm1 for all candidates at a disk rate of 9000 rpm. y-Axis error bars are for standard error of PDC fit, and x-axis error bars are for internal error of DSC. (b) Comparison of monomer remaining at week 4 in accelerated stability results, determined by SE-HPLC and the PDC as determined with custom shear device method for all candidates at a disk rate of 9000 rpm. x-Axis error bars are for standard error of PDC fit.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263191&req=5

fig07: (a) Comparison of PDC with Tm1 for all candidates at a disk rate of 9000 rpm. y-Axis error bars are for standard error of PDC fit, and x-axis error bars are for internal error of DSC. (b) Comparison of monomer remaining at week 4 in accelerated stability results, determined by SE-HPLC and the PDC as determined with custom shear device method for all candidates at a disk rate of 9000 rpm. x-Axis error bars are for standard error of PDC fit.
Mentions: The correlation of the thermal stability measured by DSC with conformational stability measured by adsorption to solid surfaces in a high-shear environment was assessed. Different antibodies are shown (Fig. 7a) in terms of their first melting temperature, Tm1, which represents unfolding of the CH2 region. This comparator is predominantly used in industrial screening of antibody candidates to determine the most stable candidates for accelerated stability studies. Comparison of the two stability indicators showed that there was no correlation between the thermal and the surface adsorption-based stability. The IgG4 candidates did show that although they were the least stable in the shear device, they had relatively good thermal stability for this transition. The lack of correlation suggests that different mechanisms of degradation are involved in the two testing methodologies.

Bottom Line: Results suggest that the techniques are orthogonal, with thermal methods based on intramolecular interaction and shear device stability based on localized unfolding revealing less stable regions that drive aggregation.Molecular modeling shows the modifications' effects on the antibody structures and indicates a possible role for Fc conformation and Fab-Fc docking in determining suspended protein stability.The data introduce the PDC value as an orthogonal stability indicator, complementary to traditional thermal methods, allowing lead antibody selection based on a more full understanding of process stability.

View Article: PubMed Central - PubMed

Affiliation: The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London, WC1E 7JE, UK; MedImmune, Granta Park, Cambridge, CB21 6GH, UK.

Show MeSH
Related in: MedlinePlus