Optimizing Hands Free Capillary Isoelectric Focusing (cIEF) (CE Pharm 2012)

In cIEF analysis, samples are pre-mixed with carrier ampholytes, pI markers, and additives. Although imaged cIEF (iCE3 IEF Analyzer) offers rapid analysis and high throughput, proteins can still experience degradation when exposed to carrier ampholytes and additives for extended periods. Automated Sample Preparation with the new iCE3 IEF Analyzer solves this problem. The system prepares the sample immediately prior to injection, limiting sample exposure to cIEF buffers and preventing degradation. There are several important method parameters such as Mixing Rate, Mixing Strokes and Mixing Depth that require optimization. This poster describes an optimization strategy for the on-board sample preparation and provides relevant examples.