Efficient Protein Purification Strategies Using Multi-Well Filter Plates

Pages: 368

Combined with chromatography media, 96-well filter plates have emerged as an efficient tool for the fractionation of small-volume protein samples. This format can be used to develop protein purification strategies and/or as a platform for moderate- to-high-throughput protein isolation. In either case, purified sample can be used for further analysis and/or downstream applications.

High-Throughput Screening on 96-Well Filter Plates

Filter plates have the capability to quickly screen various conditions. The analyst can quickly optimize the chromatography resin and purification conditions with minimal sample consumption. Once the selected sorbent is equilibrated and resuspended as a 50% slurry in the equilibration buffer, the desired amount is dispensed into the wells to a final volume of 50 μL per well. After the slurry has been dispensed into the plate’s wells, the equilibration buffer is aspirated using a multi-well plate vacuum manifold. Next, a sequence mimicking a chromatographic run is performed on the plates. For each sequence step, the corresponding solution is pipetted into the wells. Once filled, the filter plate is covered with sealing tape and incubated while shaken. The liquid is then drained from the wells using the vacuum manifold and collected in a 96-well receiver plate. The individual fractions are then analyzed by HPLC, ELISA, or other analytical methods. The flexible 96-well plate format can be used with liquid-handling robotoc systems or manual multi-channel pipettes.

Membrane-Based Protein Purification

If resin packing is inappropriate for your application, consider using ion exchange on 96-well filter plates with Mustang IEX membranes. To overcome mass transfer limitations associated with conventional resin-based chromatography, membrane chromatography was developed to obtain better flow distribution, fast flow rates, and increased accessibility for rapid purification of target proteins or removal of contaminants.