Computer-aided Assay Development for Charge Heterogeneity Analysis by iCE

Analytical techniques like isoelectric focusing have become indispensable tools in evaluating biologic preparations. The resulting surge in charge isoform analysis has led to major advances in instrumentation, such as Imaged Capillary Electrophoresis iCE. To maximize the benefits from improved instrumentation requires development of robust assays. Initially implemented in biopharmaceutical manufacturing, Quality by Design (QbD) has the potential to transform assay development. Key to QbD is comprehensively gauging the effects of process inputs on critical to quality (CTQ) attributes of the output. To this end, the Design of Experiments (DOE) methodology has proven itself to be a highly efficient tool in modeling the relationship between input and output. The contents of this poster demonstrate the successful implementation of DOE tools in the iCE3 assay development process. Utilizing a Central Composite Design (CCD) strategy, a response surface of the relationship between ampholyte composition and focusing time on peak resolution was generated. Experimental validation of this model at optimal operational settings indicated a high level of accuracy with error between the predicted and experimentally derived values ranging between -0.68 and 8.06 percent.