ANYL 126 |
| Analysis of antibody charge variants using isoelectric focusing (IEF) gels is a standard method but is semi-quantitive. Moving to capillary isoelectric focusing (cIEF) provides a method that is suitable for quantitation and validatable according to ICH requirements for the determination of charge variants. In our work, the first step was to evaluate Beckman Neutral and Agilent µSIL-FC capillaries. The Beckman and Agilent capillaries were not significantly different from each other except in the current profile. Next we validated the cIEF method using the Beckman Neutral capillary. Specificity, repeatability, intermediate precision, accuracy, range, and linearity met pre-determined validation acceptance criteria. Robustness of the method was also demonstrated. The method Quantitation Limit (QL) was determined to be 2.9% and the range for quantiation of charge species was from 2.9%-97.1%. The method is suitable for use reporting the product related substance, percent acidic species. The impact of the most critical method variables will be discussed. Other learnings related to method transfer and implementation will be discussed. |
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General Papers
7:00 PM-9:00 PM, Sunday, 10 September 2006 Moscone Center -- Hall D, Poster
Division of Analytical Chemistry |