Accelerating Protein Purification and Analysis
“Gel-in-a-shell technology” – Pall’s patented ceramic HyperD® ion exchange chromatography resin provides rapid protein purification with high capacity and good resolution.
Rapid and efficient – Obtain high speed runs (1-4 mL/min) with very little loss in binding capacity and resolution allowing more samples to be processed per day.
Higher resolution for 1 mL columns – Distinct separation of proteins for better purification.
Versatile – Luer lock inlet and outlet for convenient use with syringe, pump, or automated chromatography system.
User-friendly column design – Color-coded and labeled by chemistry type. Collar is hexagonal so columns will not unexpectedly roll off lab surface.
Instructions for Use: AcroSep™ Columns for Ion Exchange Chromatography
Methodology
The AcroSep 1 mL chromatography columns are pre-packed with Pall chromatography media for fast, convenient protein separations. The column, which uses a luer lock connector design, is compatible with a syringe, peristaltic pump, or chromatography instrument (e.g., AKTA* Explorer).
The columns take advantage of the unique, patented characteristics of our HyperD “gel-in-a-shell” resin. This design gives the high capacity of soft gel in a rigid "shell" without the problems of compressability. This resin provides reproducible packing characteristics while allowing for high binding capacities at fast flow rates.
1. Choose appropriate column chemistry.
2. Choose complimentary equilibration, loading, and elution buffers.
3. Attach column to pre-primed system (e.g., syringe, system, pump).
4. Wash and equilibrate column.
5. Add sample.
6. Wash column.
7. Elute protein with appropriate elution buffer.
8. Clean, store, and reuse column.
- Screen multiple IEX chemistries for use in your protein purification.
- Ideal for optimization studies of protein purification schemes using small sample volumes prior to scale-up.
- Efficient and reliable small scale purification of proteins for structural, functional, and yield analysis.
Materials of ConstructionColumn Housing, Cap, Plug, and Adapter: Polypropylene
Column Frit: Polyethylene
|
Media |
Function |
Color Code |
Particle Size |
Working pH |
Cleaning pH |
Ion Exchange
Capacity (1) |
|
|
|
|
|
|
|
CM Ceramic
HyperD F |
Weak cation
exchanger |
Green |
50 µm (avg) |
2-12 |
1-14 |
> 60 mg/mL (2) |
|
|
|
|
|
|
|
DEAE Ceramic
HyperD F |
Weak anion
exchanger |
Orange |
50 µm (avg) |
2-12 |
1-14 |
> 85 mg/mL (3) |
|
|
|
|
|
|
|
Q Ceramic
HyperD F |
Strong anion
exchanger |
Red |
50 µm (avg) |
2-12 |
1-14 |
> 85 mg/mL (3) |
|
|
|
|
|
|
|
S Ceramic
HyperD F |
Strong cation
exchanger |
Blue |
50 µm (avg) |
2-12 |
1-14 |
> 75 mg/mL (4) |
|
(1) Dynamic binding capacity determined at 10% breakthrough, 200 cm/h with 1.66 mL sorbent packed in a column of 5 mm ID and 100 mm height using the following:
(2) 5 mg/mL human IgG in 50 mM sodium acetate buffer, 100 mM NaCl, pH 4.7
(3) 5 mg/mL BSA in 50 mM Tris-HCl buffer, pH 8.6
(4) 5 mg/mL lysozyme in 50 mM sodium acetate buffer, pH 4.5
Column Geometry
Column Volume: 1.04 mL
Bed Height: 1.48 cm (0.58 in.)
Bed Diameter: 0.94 cm (0.37 in.)
Device Dimensions
Diameter: 1.6 cm (0.6 in.)
Length (Without Plugs): 4.8 cm (1.9 in.)
Connections
Inlet: Threaded female luer lock
Outlet: Rotating male luer locking hub
Flow Rates
Recommended: 1-4 mL/min
Back Pressure
Maximum: 3 bar (300 kPa, 43.5 psi)
Storage
2-8 ºC
The graph below illustrates dynamic binding capacity for each of our four ion exchange chemistries when run at 3.56 mL/min.
1 mL AcroSep™ Column Average Dynamic Binding Capacity (10% Breakthrough)
Data shows higher dynamic binding capacities of Pall AcroSep columns over competitive columns at higher flow rates. Each average is derived from three replicates of > 8 AcroSep columns or three replicates of two competitive columns. The proteins were at a concentration of 5 mg/mL and the choices were BSA for Q and DEAE, Lysozyme for S, and IgG for CM.
The chromatogram below is an illustration of resolution between BSA and Conalbumin using Pall's AcroSep Q column and a competitive Q column. This separation was run at 1 mL/min.
Resolution Comparison of AcroSep Columns with Q HyperD F and Competitive Q at Flow Rate of 1 mL/min
Resolution of the AcroSep columns is clearly illustrated in this chromatogram. The competitive column is represented by the red line.
The next chromatogram is an illustration of the resolution of the AcroSep Q column and the competitive column when the flow rate is elevated to 4 mL/min. It demonstrates that at 4 mL/min there is little if any change in protein separation on the AcroSep column, while the competitor performance degrades significantly with increased flow rate.
Resolution Comparison of AcroSep Columns with Q HyperD F and Competitive Q at Flow Rate of 4 mL/min
Rs = (VR2 - VR1) / [2(Wh2 - Wh1)], Rs = Resolution, VR1 = Retention of Protein 1,
VR2 = Retention of Protein 2, Wh1 = Peak width at half height of Protein 1,
Wh2 = Peak width at half height Protein 2
- AcroPrep™ Filter Plates
- Pall Chromatography Resin
- Centrifugal Devices
- Nanosep® MF (microfiltration) Centrifugal Devices
- BioTrace™, Biodyne® and FluoroTrans® Transfer Membranes
- BTS-SP Membrane
- Additional AcroSep Columns
Contact Customer Service at 800-521-1520 for possible lead-time and date item will be available to ship.
