BioSepra™ Chromatography Sorbents Offer Superior Protein Purification

Chromatography is an important technique used for the purification of individual proteins as well as the separation of proteins by class. Although chromatography was traditionally seen in protein and analytical labs, protein separations are no longer limited to protein “experts,” but are now often seen as part of proteomic, molecular biology, cell biology and biology research of all types.

 

Pall Corporation offers a full portfolio of chromatography products to meet your specific purification needs. The Pall portfolio includes both sorbent- and membrane-based chromatography tools. Pall’s portfolio features an array of products covering the most common and some very unique modes of chromatography.

 

The BioSepra chromatography sorbents can be used in traditional chromatography columns or, for quick separations, can be used in centrifugal spin columns (e.g., Pall Nanosep® devices). For high throughput separations or scouting assays, the sorbents can be combined with the AcroPrep™ filter plates with up to 96 mini-columns that can be simultaneously processed.

When performing chromatography-based purification, the choice of sorbent is crucial for ensuring effective capture (binding selectivity/capacity) and sufficient separation (resolution). Pall offers a variety of base materials ranging from soft beads (Trisacryl®, Ultrogel®, Hypercel™) to rigid beads, to a unique hybrid bead (HyperD®). Pall’s Biosepra HyperD chromatography sorbents are created using the patented “gel-in-a-shell” technology. This gel-in-a-shell bead is constructed of a high-capacity hydrogel polymerized within the gigapores of a rigid ceramic bead. The capture chemistry or ligand is attached to the hydrogel within the bead pores. The advantages of this structure are many, including higher than usual capacity, tolerance to fast chromatography runs without a concomitant increase in backpressure, and greater salt tolerance for ion exhangers.

Gel-in-a-Shell Design

The biochemical nature of the protein of interest will determine the best mode of chromatography for optimal capture/fractionation. Often, several chromatographic steps will be needed to separate or fractionate your desired protein to the level needed for your planned experimental studies.

Common modes of chromatography include ion exchange (separation by charge), size exclusion/gel filtration (separation by size), and affinity (separation using a specific capture ligand). Pall offers sorbents for all of the aforementioned modes. In addition, Pall offers unique mixed-mode chromatography sorbents including MEP Hypercel and SDR HyperD.

This chart highlights a sampling of the Pall sorbent offering:

Chromatography Method	Pall Product
Affinity	BioSepra Blue Trisacryl BioSepra Protein A Ceramic HyperD BioSepra Heparin HyperD BioSepra Lysine HyperD
Ion Exchange	BioSepra Q Ceramic HyperD BioSepra S Ceramic HyperD BioSepra DEAE Ceramic HyperD BioSepra CM Ceramic HyperD AcroPrep™ Filter Plate with Mustang™ Q Membrane AcroPrep Filter Plate with Mustang S Membrane Acrodisc® Units with Mustang Q Membrane Acrodisc Units with Mustang S Membrane
Size Exclusion	BioSepra Ultrogel AcA BioSepra Trisacryl GF Omega™ MWCO UF Membrane Centrifugal Devices AcroPrep Filter Plates with UF Membrane Minimate™ TFF Capsules
Hydroxyapatite	HA Ultrogel
Hydrophobic Charge Induction (HIC)	BioSepra MEP Hypercel

The MEP Hypercel is a unique mixed-mode sorbent for the purification of antibodies from cell culture supernatants. MEP is a synthetic, stable immunoglobulin-selective ligand that binds antibodies from all species and of all isotypes (capacity will vary). Thus, MEP offers an alternative to Protein A and Protein G for the purification of antibody and Fc fusion proteins. Additionally, antibody elution from MEP Hypercel is milder then typically used for Protein A/G and there is no concern about the downstream effect of leached Protein A in the final product. This chemistry is also quite useful for hydrophobic interaction chromatography, a hydrophobic mode without organic solvents which is much more likely to retain native structure than traditional reverse phase chromatography. The unique character of MEP HyperCel allows our customers to achieve separations that are unlike any other.

Picture of MEP Chemistry

Detergent removal is important for many applications including proteomic analysis and bioassays. SDR HyperD has been found to be ideal for detergent removal – removing Triton X100, SDS, ASB-14 and CHAPS – and generally outperforming the competition for this application. SDR HyperD can be used in rapid single-step formats or in a packed column for very large volumes. In all cases, you can feel confident that it will remove the detergent and the interference detergents for your downstream protein applications.

Pall offers the BioSepra line of chromatography sorbents bottled in bulk (1 to 100 mL) quantities. Pall also offers chromatography-based kits for abundant protein removal. These ready-to-use kits include all components necessary for depletion or selection of albumin and/or IgG from multiple species.