CHROMATOGRAPHY

Many tools and techniques are available today for protein purification. Development of a purification process requires an analysis of various strategies to isolate a protein of interest. Protein purification from a complex sample, such as cell culture or serum, requires more than one chromatographic step. Typically the first step is ion exchange (IEX) or mixed-mode chromatography to fractionate the sample and yield a subpopulation of proteins containing the target molecule. This is followed by subsequent chromatographic steps to further fractionate the sub-population until a desired purity level is reached. These steps can include affinity purification and size exclusion chromatography, as well as IEX or mixed-mode chromatography to isolate the target protein.

Each step in the purification process requires optimization in order to maximize yield and purity of the target protein. Thus, small-scale purification experiments are used during the development and optimization phase to preserve precious sample while providing crucial information. The methods demonstrate the use of AcroPrep™ Advance 96-well filter plates combined with Pall chromatography media as an efficient tool for fractionation of small-volume protein samples. The format can be used to develop protein purification strategies and/or as a platform for moderate to high throughput needs.

AcroPrep Advance 96-well filter plates offer improved consistency and reproducibility;

• Well design increases well-to-well consistency for sample processing and recovery.
• Outlet tips minimize the formation of hanging drops, reducing the chance of cross contamination following filtration.
• Optimized membrane configurations for assays, such as multiplexing.
• Individually sealed membranes and biologically inert housing minimizes the possibility of sample cross-contamination or loss.
• Designed to meet SBS/ANSI guidelines, ensuring compatibility with standard processing equipment.
• Smooth top surface, textured window for labelling, and A1 corner notch allow for easy identification.