Chromatography

Affinity Chromatography

Affinity chromatography separates compounds based on the properties of chosen ligands with specific affinity to the molecule being selected for attachment to the solid matrices. Often, the binding between the molecule and the ligand can be reversed after the selection step. This is done, for example, through buffer, salt concentration, or pH change.

Pall offers bottled sorbents, packaged kits, and pre-packed columns for affinity chromatography. Pall bottled sorbents can be used for small and large sample sizes, and for single use and high throughput methods of purification. Our base sorbent varies depending on targeted application.

Protein A Ceramic HyperD® F Sorbent is a high capacity affinity sorbent designed for process-scale purification of immunoglobulin G. The sorbent combines ease of use with high binding capacity and excellent scalability. Protein A Ceramic HyperD F sorbent is prepared using a rigid proprietary ceramic bead. Recombinant Protein A is immobilized to a specially formulated hydrogel within the porous ceramic bead.

Blue Trisacryl® M Sorbent is an affinity chromatography sorbent used for the purification of a wide variety of enzymes and proteins such as kinases, albumin, interferons, and some coagulation factors. The base matrix is Trisacryl GF2000, a macroporous non-ionic sorbent on which Cibacrons Blue is covalently immobilized. Cibacron Blue F3GA dye is strongly bound to the matrix through a six carbon spacer arm. The reaction is performed with the EEDQ coupling agent. This type of coupling prevents any leakage of the dye in normal working conditions. (See page 49.)

Heparin HyperD M Sorbent is a high capacity affinity preparative sorbent for the purification of biological molecules that bind to heparin, such as coagulation factors, growth factors, lipoproteins, etc. Heparin HyperD M affinity sorbent employs a high-capacity hydrogel polymerized within the large pores of a rigid bead. This design combines the desirable characteristics of a soft, high capacity hydrogel with the high dimensional stability of a rigid bead.

Lysine HyperD Sorbent is a high capacity affinity preparative sorbent for the purification of biological molecules that bind to lysine such as plasminogen from human or animal species plasma. Lysine HyperD affinity sorbent employs a highcapacity hydrogel polymerized within the large pores of a rigid bead. This design combines the desirable characteristics of a soft, high capacity hydrogel with the high dimensional stability of a rigid bead.

Type of Affinity Purification	Ligand(s)	Pall Product
Albumin Depletion	Blue Dye	Blue Trisacryl M Chromatography Sorbent
IgG Purification	Protein A MEP	Protein A Ceramic HyperD Chromatography Sorbent, MEP HyperCel™ Chromatography Sorbent
Lipoprotein Purification	Heparin	Heparin HyperD M Chromatography Sorbent
Glycoprotein Purification	Lysine Hydroxyapatite	Lysine HyperD Chromatography Sorbent HA Ultrogel® Chromatography Sorbent

Metal Chelate Affinity Chromatography

Metal chelate affinity chromatography exploits the affinity of proteins or other molecules for metal ions. The affinity is derived from the formation of coordination bonds between the metal ions and certain exposed side chains of protein amino acids. Histidine, present in many proteins, forms complexes with transition metal ions such as Cu^–, Zn²⁺, Ni²⁺ or Fe³⁺.

IMAC HyperCel Sorbent uses tridentate IDA (iminodiacetic acid) as a chelating ligand agent. This ligand is immobilized on the HyperCel base sorbent, a stable, robust, and wellknown sorbent used for both research and industrial-scale protein separations.

Summary of Pre-Fractionation Options for IMAC HyperCel Sorbent

*Elution may require several steps, such as imidazole, pH, salt, EDTA, or EGTA and detergents linked together to achieve efficient recovery of bound material.

Ion Exchange Chromatography

Ion exchange chromatography separates compounds based on net surface charge. Molecules are classified as either anions (having a negative charge) or cations (having a positive charge). Some molecules (e.g., proteins) may have both an anionic and cationic group. A positively-charged support (anion exchanger) will bind a compound with an overall negative charge. Conversely, a negatively-charged support (cation exchanger) will bind a compound with an overall positive charge. Ion exchange matrices can be further categorized as either strong or weak. Strong ion exchange matrices are charged (ionized) across a wide range of pH levels. Weak ion exchange matrices are ionized within a narrower pH range. The four most common ion exchange chemistries are shown below.

Type of Ion Exchanger	Common Abbreviation	Functional Group	Pall Product
Strong Anion	Q	Quarternary Ammonium	Q HyperCel™ Acrodisc® with Mustang® Q Q Ceramic HyperD®F AcroPrep™ Advance with Mustang Q AcroSep™ with Q Ceramic HyperD®F
Weak Anion	DEAE	Diethylaminoethyl	DEAE Ceramic HyperD®F AcroSep with DEAE Ceramic HyperD®F
Strong Cation	S	Sulfonic Acid	S HyperCel Acrodisc with Mustang S S Ceramic HyperD®F AcroPrep Advance with Mustang S AcroSep with S Ceramic HyperD®F
Weak Cation	CM	Carboxymethyl	CM Ceramic HyperD®F AcroSep with CM Ceramic HyperD®F
Weak Anion	Not Relevant	Primary amine	HyperCel STAR AX AcroPrep™ ScreenExpert Plate filled with HyperCel STAR AX

Pall offers ion exchange sorbents, pre-packed columns, and membranes. In many areas, chromatography sorbents are the media of choice for chromatography applications, but in some cases where sorbent-based methods have limitations (e.g., purification of viruses or large molecules) membranes have proven to be a robust, scalable, and economic alternative. Membranes perform well in such applications because of their faster flow rates compared to sorbents.

What Flow Rate to Use?

Mustang membrane chromatography devices are designed to run at flow rates of at least 10 column volumes per minute. Initial optimization of buffer selection, pH, capacity, and elution conditions can all be performed at this flow rate. Faster flow rates for equilibration, loading, and washing will give better throughput; a slower flow rate during binding and elution may give better resolution for some processes. The open structure of Mustang membranes does not require diffusion into pores, and therefore normally permits high flow rates.

What Buffer to Use?

Typically, ion exchange chromatography matrices are loaded in low ionic strength buffers. Under these conditions, charged macromolecules will be retained by the stationary phase bearing the opposite charge. Macromolecules bearing the same charge as the stationary phase will simply flow through without binding. The ion exchange matrix is washed with additional low ionic strength buffer to completely wash out any remaining unbound species, and the bound species are differentially eluted by buffers containing increasing amounts of salt. As the ionic strength of the mobile phase increases, salt ions compete for binding to the charges on the ion exchange matrix, displacing the bound macromolecule, and allowing them to elute from the matrix. To avoid difficulty, use anionic (negatively-charged) buffers for cation exchange, and cationic (positively-charged) buffers for anion exchange.

What pH to Use?

Although pH does not influence the charge of a strong ion exchange matrix, it will influence the charge on the macromolecules in solution. The operating pH in ion exchange chromatography is selected to maximize the resolution of the target molecule from the contaminant background. In some cases, a pH is selected to provide maximum binding of the target molecule and minimum binding of the contaminants (positive mode). Elution of the target molecule is accomplished by increasing the salt concentration. In other cases, a pH is selected to provide maximum binding of the contaminants and minimal or no binding of the target molecule (negative mode). The target molecule ends up in the flow through, and the contaminants are separated away by binding to the matrix. Through careful selection of both the ion exchange matrix and the operating pH, both yield and purity can be maximized in a single step. However, it is never possible to achieve 100% purity in a single step, which is why multiple steps must be sequenced together to take advantage of the variety of chemical differences between the target molecule and background contaminants.

What Salt to Use for Elution?

After binding, salt concentrations for elution are chosen so the target molecule does not co-elute with contaminants that have also bound to the ion exchange matrix. Ions of the eluting salt must displace other molecules from the charged groups on the stationary phase with either a gradient or step in the 0 to 1.0 M range. The effectiveness of displacement for commonly used cations is: Ca²⁺ > Mg²⁺ > Na⁺ > K⁺ > NH₄ ⁺.

The order of displacement effectiveness for commonly-used anions is: PO₄ ^3- > SO₄ ^2- > COO^- > Cl^-. These rankings correlate with the Hofmiester series, and the strongest eluting salt is not always best. Ideally, several salts should be tested, and finding optimum elution conditions often involves trial and error. Most users will start with either NaCl or KCl simply because they are readily available in the lab. However, CaCl2 or MgCl2 may be used. For some proteins, those salts may actually end up being a better choice. Regardless of eluting salt selection, the effect on the purity, stability, and activity of the target molecule will have to be assessed.

Gel-in-a-Shell Technology: The Enhanced Diffusion Concept

Traditional macroporous ion exchangers operate on the basis of classical pore diffusion. Pore diffusion is characterized by rapidly decreasing binding capacity with increasing flow rate. In contrast, the unique structure of Pall's Ceramic HyperD® sorbent supports a more rapid mechanism of mass transfer, known as enhanced diffusion. Rapid mass transfer overcomes classical flow rate dependence. Because product is bound throughout the gel-filled pore (not merely at the interior surface of the pore), total binding capacity is enhanced.

Using enhanced diffusion, Pall's gel-in-a-shell technology binds product throughout the gel-filled pore, enhancing total binding capacity.

Binding of protein within the hydrogel carries an extraordinarily high concentration of ion exchange functional groups: 150 - 400 μeq/mL. The average distance between charged sites on the hydrogel is ~20 Å. Thus, a protein molecule within the gel is simultaneously in contact with a large number of ion exchange sites. It remains in contact with a similar number of sites no matter where it moves within the threedimensional structure of the hydrogel. As a result, the protein is energetically unconstrained and may migrate freely. Protein diffuses rapidly within the hydrogel to give a homogeneous distribution, facilitating uptake of additional material from the solution. Under binding conditions, strong attractive electrostatic forces between the highly substituted hydrogel and the protein drive entry of protein into the gel. The table below demonstrates the high dynamic binding capacity of various Ceramic HyperD sorbents.

Ceramic HyperD F and HyperCel STAR AX sorbents are available in bulk as bottled sorbents, or pre-packed into Pall's AcroSep™ chromatography columns.

Pall Ion Exchange Sorbents Exhibit High Dynamic Binding Capacity While Maintaining Fast Flow Rates ; HyperCel STAR AX sorbent has a differentiated anion exchange selectivity, and unique salt tolerance

Media	Flow Rate (mL/min)
	1	5	10
DEAE Ceramic HyperD F	101.5 mg/mL	87.5 mg/mL	77.5 mg/mL
S Ceramic HyperD F	80.5 mg/mL	61.5 mg/mL	53.5 mg/mL
CM Ceramic HyperD F	108.0 mg/mL	87.5 mg/mL	73.5 mg/mL
Q Ceramic HyperD F	Not available	Not available	Not available

Ceramic HyperD F ion exchange sorbents have high dynamic binding capacity (50-110 mg BSA/mL) . Dynamic binding capacity is measured in a 1 mL packed column by pumping BSA (anion) or lysozyme (cation) at 5 mg/mL in binding buffer until the column capacity is exceeded. The capacity is then calculated by estimating the volume of protein required to achieve this "breakthrough" and expressed as mg/mL sorbent volume. Q/S/ HyperCel and HyperCel STAR AX sorbents are based on the robust HyperCel matrix ; HyperCel STAR AX brings differentiated selectivity (primary amine weak anion exchange ligand), and bind proteins and contaminants in a broad feedstock conductivity range (2-15 mS/cm), limiting unit operations such as dilution or UF/DF.

Using Mustang® Membranes for Ion Exchange Chromatography

Membranes are recommended in chromatography applications when there is a need to purify large molecules or in situations where faster flow is needed. Membrane chromatography is extremely economical because flow rates are significantly faster than traditional sorbent chromatography, decreasing cycle time and increasing throughput. Pall's Mustang membranes possess large convective pores and have dynamic binding capacities that are relatively insensitive to the effects of high flow rates, even for large molecules such as plasmids and viruses. Pall's membrane devices for ion exchange chromatography offer:

• True scalability – For laboratory-scale applications, Mustang membranes are available in Acrodisc® units for single samples and AcroPrep™ Advance filter plates for higher sample processing. Devices with Mustang Q and S membranes can be scaled up to larger-capacity capsules and cartridges from Pall.
• Application-specific membrane chemistries – Mustang Q membrane is a strong anion exchanger that effectively binds plasmid DNA, negatively-charged proteins, and viral particles. Mustang S membrane is a strong cation exchanger that effectively binds positively-charged proteins and viral particles.
• High binding capacities and fast flow rates – Mustang membranes withstand high flow rates to render faster purification without affecting recovery rates.

Mustang Q Membrane for Anion Exchange

Mustang Q membrane is an anion exchanger with a polyethersulfone (PES) base modified with quaternary amines. Mustang Q membrane delivers efficient and rapid flow rates with a convective pore structure combined with high dynamic binding capacity for plasmid DNA (3.6 mg/Acrodisc unit), negatively-charged proteins (10 mg), and viruses (1012 viral particles). Processing time is much shorter and more efficient than the conventional bead- or sorbent-based technology. Mustang devices have throughputs of up to 100 times that of traditional columns, with no associated loss of binding capacity. The syringe filter and multi-well plate formats can directly scale up to larger-scale capsules and cartridges for larger-volume applications.

Mustang S Membrane for Cation Exchange

High capacity Mustang S membrane is a cation exchanger with a polyethersulfone (PES) base modified with sulfonic acid groups. Mustang S membrane delivers efficient and rapid flow rates with a convective pore structure combined with high dynamic binding capacity for positively-charged proteins and viruses. Processing time is much shorter and more efficient than the conventional bead or sorbent-based technology. The syringe filter and multi-well plate formats can directly scale up to larger-scale capsules and cartridges for larger-volume applications.

Mustang membrane is available in Acrodisc units for single sample processing.

Mixed-Mode Chromatography

Hydrophobic Charge Interaction Chromatography (HCIC)

HCIC is based on the pH-dependent behavior of ionizable, dual-mode ligands. Adsorption is based on mild hydrophobic interaction and is achieved without addition of lyotropic or other salts. Desorption is based on charge repulsion. It is performed by reducing pH.

MEP HyperCel™ sorbent is a high capacity, high selectivity sorbent designed for the capture and purification of monoclonal and polyclonal antibodies. HCIC is based on the pH dependent behavior of ionizable, dual-mode ligands. MEP HyperCel sorbent carries an antibody-selective ligand, 4-Mercapto-Ethyl-Pyridine (4-MEP). Adsorption is based on mild hydrophobic interaction and is achieved without addition of lyotropic or other salts. Desorption is based on charge repulsion. It is performed by reducing the pH.

HEA and PPA HyperCel sorbents are novel industry scalable chromatography sorbents designed for protein capture and impurity removal in a biopharmaceutical environment. Operating on a "mixed-mode" mechanism, their chromatographic behavior is based on a combination of electrostatic and hydrophobic properties of the protein and ligands. HEA and PPA HyperCel sorbents provide unique and different selectivities not accessible with traditional ion exchange or HIC, that can be screened to facilitate process development. For example, the mixed-mode interaction mechanism can be exploited to achieve discrimination of proteins having similar or very close isoelectric points, a separation which cannot be performed by methods like ion exchange.

Hydroxyapatite Chromatography

Hydroxyapatite chromatography is considered to be a "pseudo-affinity" chromatography, or "mixed-mode", ion exchange. It has proven to be an effective purification mechanism in a variety of processes, providing biomolecule selectivity complementary to more traditional ion exchange or hydrophobic interaction techniques. HA Ultrogel® is easily scalable and is currently used in research scale to multi-liter column applications.

HA Ultrogel sorbent available from Pall is a hydroxyapatite agarose composite sorbent for the separation of biomolecules from research and development scale to manufacturing. HA Ultrogel is a cross-linked tri-dimensional composite based on spherical agarose beads with entrapped microcrystals of hydroxyapatite.

Detergent-Removal Chromatography

Plasma preparations may contain viruses that are effectively removed and inactivated by combining nanofiltration (for size exclusion removal of non-lipid enveloped viruses) and treatment with non-ionic solvents and detergents (effective for lipid-coated viruses). The elimination of solvent and detergent from biological fractions is necessary, and can be achieved by various methods including sorbent partitioning, size exclusion, affinity, or batch extraction with vegetable oils combined with reverse phase chromatography on C18.

SDR HyperD® sorbent is a unique sorbent designed to eliminate solvent and detergent from biological fluids. SDR HyperD sorbent is made of silica beads in which the pore volume is filled with a three-dimensional cross-linked hydrophobic polymer. The particle size distribution (40 - 100 μm), the small pore size of the silica beads, and the hydrophobic nature of the chemical groups make SDR HyperD sorbent an excellent tool for specific solvent and detergent removal from biological liquids.

Bulk chromatography sorbents are available in various packaging configurations for use in differently-sized chromatography columns.

Desalting Applications

Specification	Ultrogel AcA 202 Sorbent
Particle Size	60 - 140 μm
Monomer	20% (w/v) acrylamide
Cross-Linker	2% (w/v) agarose
Exclusion Limit	22,000
Linear Fractionation Range	1,000 - 15,000
Resolving Power (plates/m)	3,000
Working pH Range	3 - 10

Size Exclusion Applications

Application	Pall Products
Nucleic Acid Concentration	AcroPrep™ Advance filter plates with ultrafiltration membranes
Nucleic Acid Desalting/ Buffer Exchange	Centrifugal devices with ultrafiltration membranes
Protein Fractionation	Ultrogel AcA sorbents
Protein Buffer Exchange	Minimate™ TFF system AcroPrep™ Advance filter plates with ultrafiltration membranes

Chromatography Columns (Pre-Packed)
AcroSep™ chromatography columns with Blue Trisacryl® M sorbent	•
AcroSep chromatography columns with Ceramic HyperD® F sorbents (Q, S, CM, DEAE)		•
AcroSep chromatography columns with HEA HyperCel™ sorbent			•
AcroSep chromatography columns with IMAC HyperCel sorbent	•
AcroSep chromatography columns with MEP HyperCel sorbent			•
AcroSep chromatography columns with PPA HyperCel sorbent			•
AcroSep chromatography columns with Protein A Ceramic HyperD F sorbent	•
AcroSep chromatography columns with SDR HyperDsorbent			•
PRC Chromatography Columns		•	•
Chromatography Sorbents
Blue Trisacryl M sorbent	•
Ceramic HyperD ion exchange sorbents (Q, S, DEAE, CM)		•
HA Ultrogel® sorbent			•
HEA HyperCel sorbent			•
Heparin HyperD M sorbent	•
IMAC HyperCel sorbent	•
Lysine HyperD sorbent	•
MEP HyperCel sorbent			•
PPA HyperCel sorbent			•
Protein A Ceramic HyperD F sorbent	•
Q and S HyperCel sorbents		•
SDR HyperD sorbent			•
Filter Plates
AcroPrep™ 384-well filter plates with GHP membrane	•	•	•	•
AcroPrep 384-well filter plates with Supor® membrane	•	•	•	•
AcroPrep Advance 96-well filter plates for protein purification	•	•	•	•
AcroPrep Advance 96-well filter plates for ultrafiltration				•
Filtration Syringe Filters
Acrodisc® PSF GxF syringe filters with GHP membrane	•	•	•	•
Acrodisc PSF syringe filters with GHP membrane	•	•	•	•
Acrodisc unit with Mustang® Q membrane		•
Acrodisc unit with Mustang S membrane		•
Hardware
Multi-well plate vacuum manifold	•	•	•	•
Vacuum/pressure pumps	•	•	•	•

Pall offers a comprehensive, versatile, and targeted range of chromatography sorbents and membranes to support your protein purifications. Our portfolio of products facilitates applications ranging from research, to process development and scale up, to full-scale process manufacturing.

Affinity and Metal Chelate Chromatography

Sorbent	Ligand	Average Particle Size (μm)	Working / Cleaning pH	Applications
Protein A Ceramic HyperD® F sorbent	Recombinant Protein A	50	2 – 11 / 2 – 13	Immunoglobulins MAbs
Blue Trisacryl M sorbent	Blue dye	60	1 – 10 / 1 – 10	Albumin, enzymes Growth hormones Growth factors Interferon Coagulation factors Lipoproteins
Heparin HyperD M sorbent	Porcine heparin	80	3 – 13 / 3 – 13	Coagulation factors Growth hormones Enzymes Lipoproteins Nucleic acid
Lysine HyperD sorbent	L-lysine	70	3 – 13 / 3 – 13	Plasminogen Glycoproteins
IMAC HyperCel™ sorbent	Imino-diacetic acid (IDA)	90	3 – 12 / 3 – 14	Immobilized metal affinity chromatography (IMAC)

Ion Exchange Chromatography

Sorbent	Chemical Group	Size (μm)	Working / Cleaning pH	Applications
Q, S HyperCel™ sorbents	Quaternary amine (Q) Sulfonic acid (S)	75	1 – 13 / 1 – 14	High productivity separation Recombinant proteins Monoclonal antibodies (MAbs) Plasma derivatives Biosimilars Vaccines Polypeptides Plasmid purification High resolution for recombinant proteins
Q, S, DEAE, CM Ceramic HyperD F sorbents	Quaternary amine (Q) Sulfopropyl (S) Diethylaminoethyl (DEAE) Carboxymethyl (CM)	50	2 – 12 / 1 – 14	Recombinant proteins Plasmid purification Proteins, vaccines MAbs Capture step Immunoglobulin purification
Membrane	Chemical Group	Typical DBC	Available Format	Applications
Mustang® Q	Quaternary amine (Q)	70 mg/mL BSA 30 mg/mL DNA	Single-use capsules, reusable XT capsules, membrane, filter plates, syringe filters	Contaminant removal in polishing applications (DNA, HCP, viruses, endotoxins) Plasmid, virus, high molecular weight protein capture, and oligonucleotide purification Column guard to enhance selectivity of subsequent chromatography steps
Mustang S	Sulfonic acid (S)	60 mg/mL human IgG	Single-use capsules, membrane, filter plates, syringe filters	Specific capture (baculoviruses) Aggregate polishing from MAb feedstream

Mixed-Mode Chromatography

Sorbent	Ligand	Average Particle Size (μm)	Working / Cleaning pH	Applications
MEP HyperCel sorbent	4-Mercaptoethyl-pyridine	90	2 – 12 / 2 – 14	Direct capture of polyclonal and monoclonal antibodies of most classes, sub-classes, and species Enzymes and recombinant proteins Capture of recombinant antibody fragments Separation of monomeric IgG from aggregates Direct capture of proteins in low salt concentration
HEA HyperCel sorbent	Hexylamine	90	2 – 12 / 1 – 14
PPA HyperCel sorbent	Phenylpropylamine	90	2 – 12 / 1 – 14
SDR HyperD sorbent	Hydrophobic polymer moiety	70	2 – 12 / 2 – 12	Elimination of solvent and detergent from biological fluids
Membrane	Nature	Typical DBC	Available Format	Applications
Mustang E	Positively-charged modified hydrophilic polyethersulfone (200 nm pore size) (E)	4 x 106 EU/mL	Single-use capsules, syringe filters, membrane	Endotoxin removal from buffers, water, saline, and process streams

Hydroxyapatite Chromatography

Sorbent	Nature	Average Particle Size (μm)	Working / Cleaning pH	Applications
HA Ultrogel® sorbent	Hydroxyapatite crystals	120	5 – 13 / 5 – 14	Immunoglobulin separation Glycoproteins, vaccines

Chromatography may be the most important separation tool in today's arsenal of protein purification techniques. Pre-packed columns and other formats of sorbent-containing devices are becoming more valuable to researchers because they save time and provide reliable, efficient results. Today, sorbent-containing technologies are available with many different sorbent types, and ion exchange is one of the most popular. Using ion exchange, proteins can be fractionated and separated very precisely. Proteins can be separated with high resolution even if there is only a small difference in charge by method optimization. Affinity chromatography involves specific interactions between biomolecules. Ligands are covalently bound to a support matrix and the biomolecules of interest will adhere to the ligands in the column, resulting in a very pure separation.

Pall is continuing to expand our line of chromatography sorbents for laboratory applications to include both bulk sorbents and pre-packed columns. Documented reliability, outstanding quality, and ease of use make Pall chromatography products an outstanding choice for protein purification applications.

User-Friendly Column Design in Multiple Chemistries

Pre-packed AcroSep columns feature convenient, colorcoded collars to help end users easily identify the sorbent type. These collars are hexagonally-shaped to prevent the columns from rolling off a table surface.

AcroSep Selection Guide and Color Coding

Type of Purification	Sorbent	1 mL AcroSep Column	Color
Ion Exchange, Weak Cation	CM Ceramic HyperD® F	AcroSep™ Ion Exchange Chromatography Columns	Green
Ion Exchange, Strong Cation	S Ceramic HyperD F	AcroSep™ Ion Exchange Chromatography Columns	Sky Blue
Ion Exchange, Strong Anion	Q Ceramic HyperD F	AcroSep™ Ion Exchange Chromatography Columns	Red
Ion Exchange, Weak Anion	DEAE Ceramic HyperD F	AcroSep™ Ion Exchange Chromatography Columns	Orange
Affinity	Protein A Ceramic HyperD F	AcroSep™ Chromatography Columns for Affinity Purification	Pearl
Affinity	IMAC HyperCel™	AcroSep™ Chromatography Columns for Affinity Purification	Cobalt Blue
Affinity	Blue Trisacryl® M	AcroSep™ Chromatography Columns for Affinity Purification	Dark Blue
Mixed-Mode	MEP HyperCel	AcroSep™ Chromatography Columns for Mixed-Mode	Purple
Mixed-Mode	HEA HyperCel	AcroSep™ Chromatography Columns for Mixed-Mode	Black
Mixed-Mode	PPA HyperCel	AcroSep™ Chromatography Columns for Mixed-Mode	Yellow
Detergent Removal	SDR HyperD	AcroSep™ Chromatography Columns with SDR HyperD® Detergent Removal Sorbent	Natural

AcroSep columns are provided with luer inlet and outlet fittings to facilitate ease of use with a syringe, pump, or chromatography system.

AcroSep ion exchange columns feature Pall's patented ceramic HyperD sorbent with "gel-in-a-shell" technologyfor rapid protein purification with high capacity and good resolution. Users can obtain high speed runs (1 - 4 mL/min) with very little loss in binding capacity and resolution, which allows more samples to be processed per day. These columns are useful for:

• Screening multiple IEX chemistries for use in protein purification.
• Optimizing studies of protein purification schemes using small sample volumes prior to scale-up.

AcroSep columns with SDR HyperD detergent removal sorbent provide high dynamic binding capacity for fast and efficient removal of many different detergents. The columns exhibit high recovery of proteins (exclusion limit 10 kDa) and high adsorption capacity for small hydrophobic molecules. The SDR sorbent is stable in acidic, polar organic, and oxidizing solutions. Use these columns for:

• Removing various (anionic, cationic, and non-ionic) detergents from proteomics samples.
• Removing detergents commonly used in membrane protein solubilization.

Continuing Advancements

AcroSep chromatography columns for affinity purification provide high dynamic binding capacity when purifying a variety of biomolecules. The enhanced coupling mechanism removes concerns of ligands leaching into purified samples. These columns are ideal for:

• IgG affinity purification from multiple sample types.
• Purification of His-tagged proteins.
• Depletion of albumin from plasma and serum samples.

AcroSep chromatography columns for mixed-mode feature Pall's HyperCel sorbent, which provides high porosity, chemical stability, and low non-specific interaction. Binding of target molecules typically occurs at physiological pH and removes the need for high salt concentration used in conventional hydrophobic interaction chromatography. Use these columns for:

• Antibody capture and purification.
• Direct hydrophobic capture and purification.
• Separation of proteins with similar isoelectric points.