HiTrap Q FF Column
Strong Anion Exchange Column for Fast Small-Scale Protein Purification
HiTrap Q FF is a strong anion exchange chromatography column designed for small-scale protein purification. Packed with Q Sepharose Fast Flow resin, it provides convenient, high-flow ion exchange separations with good scale-up potential.
Product Overview
HiTrap Q FF is a strong anion exchange chromatography column for small-scale protein purification. It is packed with Q Sepharose Fast Flow strong anion exchange resin.
The column format is convenient and affordable for fast ion exchange separations, either as a single column or connected in series. HiTrap Q FF provides high flow rates and good scale-up potential.
The resin consists of 6% cross-linked agarose with an average particle size of approximately 90 μm. The HiTrap format allows convenient connection to a syringe, peristaltic pump, or chromatography system.
Key Features
Strong Anion Exchange
Functionalized with a strong anion exchange group for purification of charged biomolecules.
Q Sepharose Fast Flow
Packed with Q Sepharose Fast Flow resin for efficient ion exchange chromatography.
90 μm Particles
Resin has an average particle size of approximately 90 μm.
Convenient HiTrap Format
Easy connection to a syringe, peristaltic pump, or chromatography system.
Broad pH Stability
Operational pH range from 2–12 and cleaning-in-place stability from pH 2–14.
Scale-Up Potential
High flow rates and convenient operation support method development and scale-up.
Applications
Method Scouting
Useful for screening ion exchange conditions during protein purification method development.
Group Separations
Designed for separation of charged biomolecules based on their interaction with the strong anion exchange resin.
Sample Concentration
Can be used as part of workflows for concentrating charged biomolecules.
Sample Clean-Up
Suitable for sample clean-up before downstream purification or analytical procedures.
Technical Specifications
| Parameter | Specification |
|---|---|
| Product | HiTrap Q FF Column |
| Cat. No. | Hi-236P |
| Ionic Exchanger Type | Strong anion |
| Functional Group | -N+(CH3)3 |
| Matrix | Cross-linked agarose, 6%, spherical |
| Average Particle Size | ~90 μm |
| Dynamic Binding Capacity | ~120 mg HSA/mL resin |
| Metal Ion Capacity | 0.18–0.24 mmol Cl-/mL resin |
| Recommended Flow Rate | <4 mL/min |
| Recommended Column Height | 25 mm |
| Dimensions | 7 × 25 mm |
| Column Volume | 1 mL |
| Column Internal Diameter | 7 mm |
| Maximum Operating Pressure | 5 bar (0.5 MPa), 70 psi |
| Column Hardware Pressure Limit | 5 bar (0.5 MPa) |
| pH Working Range | 2–12 |
| CIP Stability | 2–14 |
| Chemical Stability | Stable to commonly used aqueous buffers, 1.0 M NaOH, 8 M urea, 6 M guanidine hydrochloride and 70% ethanol. |
| Storage | 4–30°C, 20% Ethanol |
| Pack Size | 5 × 1 mL |
Sample Preparation
The sample should be adjusted to the composition of the start buffer by buffer exchange before loading onto the column.
Suitable options for buffer exchange include HiTrap Desalting, HiPrep 26/10 Desalting or PD-10 columns.
Purification Procedure
Fill with Start Buffer
Fill the syringe or pump tubing with start buffer at low ionic strength.
Connect the Column
Remove the stopper and connect the column to the syringe using the provided connector, or to the pump tubing, drop to drop, to avoid introducing air.
Remove Outlet End
Remove the snap-off end at the column outlet.
Wash Preservatives
Wash out preservatives with 5 column volumes of start buffer. Use 1 mL/min for HiTrap 1 mL and 5 mL/min for HiTrap 5 mL.
Wash with Elution Buffer
Wash with 5 column volumes of elution buffer consisting of start buffer with 1 M NaCl.
Equilibrate
Finally equilibrate the column with 5–10 column volumes of start buffer.
Apply Sample
Apply the sample at 1 mL/min for HiTrap 1 mL and 5 mL/min for HiTrap 5 mL using a syringe fitted to the Luer connector or by pumping it onto the column.
Wash the Column
Wash with at least 5 column volumes of start buffer or until no material appears in the eluate.
Elute
Elute with 5–10 column volumes of elution buffer according to the selected gradient type.
Desalt if Necessary
Purified fractions can be desalted using HiTrap Desalting, HiPrep 26/10 Desalting or a PD-10 column if necessary.
Regenerate
After elution, wash with 5 column volumes of regeneration buffer consisting of start buffer with 1 M NaCl.
Re-equilibrate
Finish with 5–10 column volumes of start buffer. The column is then ready for a new sample.
Buffer & Operating Conditions
Start Buffer
Low ionic strength buffer used for column equilibration and sample loading.
Elution / Regeneration Buffer
Start buffer containing 1 M NaCl.
Chemical Stability
The resin is stable to commonly used aqueous buffers, 1.0 M NaOH, 8 M urea, 6 M guanidine hydrochloride, and 70% ethanol.
Frequently Asked Questions
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HiTrap Q FF Column
Strong anion exchange chromatography for efficient protein purification, method scouting, sample clean-up and charged biomolecule separation.