HPLC

DiscoveryBIO PolyMA WAX

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Ordering Information

Discovery BIO PolyMA-WAX
Polymethacrylate polymer-based weak anion-exchange HPLC column
 
Discovery BIO PolyMA-WAX polymer-based weak anion-exchange particles have discriminating hydrophilic surface chemistry making them ideally suited for separating proteins, peptides, and other biotechnology-derived products.

Choose Discovery BIO PolyMA-WAX for anion-exchange separations. Generally run at a pH greater than the protein's pI (isoelectric point) - usually pH 7 or higher.
 
Particle:   Polymethacrylate with hydrophilic coating
Functional Group:   Diethylaminoethyl (DEAE)
Counterion (as supplied):   Cl-
Particle Shape:   Spherical, monodispersed
Particle Size:   5µm
Pore Size:   1000Å
Coverage:   0.3meq/g
pH Range:   2 to 10*
Temperature Range:   4°C to 50°C
 
* PolyMA-WAX is a weak anion-exchange material. It can be used at high pH values but with reduced charge.
 
Solutions for Protein and Peptide Separation Challenges
 

Excellent separations of protein isoforms

High resolution at low sample load

Quantitative recovery - a hydrophilic surface eliminates protein adsorption

High efficiency

Wide pH range


Here's the
Proof
 
EFFICIENT ANION-EXCHANGE SEPARATIONS
 
Discovery BIO PolyMA-WAX typically has higher efficiency than competitive polymeric ion-exchange materials.
 
Comparison of Efficiency: Hemoglobin Variants
Columns:   Discovery BIO PolyMA-WAX, 5cm x 4.6mm, 5µm (Cat. No. 59602-U) and Competitive polymeric weak anion exchange columns
Mobile Phase:   (A) 10mM Tris/HOAc, pH 8.0;
(B) 10mM Tris/HOAc, 0.25M KCl, pH 8.0
Flow:   3.01cm/min (flow rates appear in Figure)
Temp.:   35°C
Detection:   UV, 280nm
Sample:   50µg each variant
Gradient Profile:   1.6% B per minute. See Figure for details.
     
1.   Hemoglobin A2
2.   Hemoglobin S
3.   Hemoglobin A0
 
 
BALANCE OF ION-EXCHANGE CAPACITY AND PROTEIN RECOVERY
 
Although capacity is dependent on the protein under study and operating conditions, generally between 20 and 50mg of total protein can be injected onto the Discovery BIO PolyMA-WAX columns.
 
Ion-exchange Capacity
Column Protein mg per mL of column volume mg per column
Discovery BIO PolyMA-WAX BSA 50mg 40mg
 
Loading studies were run at 240cm/hr (0.67mL/min) using 0-1M NaCl salt gradients in 20mM Tris-HCl, pH 8. Protein concentration was 2mg/mL in starting mobile phase.
 
Protein (Fibrinogen) Recovery
  Fibrinogen Recovery
Column 1st Injection 2nd Injection 5th Injection
Discovery BIO PolyMA-WAX (5µm, 5cm x 4.6mm) 88.9% 89.9% 92.9%
Competitive polymethacrylic DEAE column (10µm, 7.5cm x 7.5mm) 59.1% 75.7% 82.1%
 
Mobile Pphase: Gradient of 0 - 0.5M NaCl in 20mM Tris-HCl (pH 8) over 30 minutes. Flow: 1mL/min. Ambient temperature. Sample: 40µg fibrinogen (Sigma part no. F-4753) in 20µL starting mobile phase.

COLUMN STABILITY AND LIFETIME
 
Column stability is important from quality and economic standpoints. Stable columns give reliable results. Stable columns cost less per injection and cause less system down-time. Results obtained on Discovery BIO PolyMA-SCX and PolyMA-WAX will be reproducible injection after injection because of exceptional column stability. In the figure below, Discovery BIO PolyMA-WAX columns are shown to give stable retention of three proteins after 500 injections, with no sign of deterioration. The same high degree of stability is shown for Discovery BIO PolyMA-SCX columns using three different proteins in the figure below.
 
Stability of Discovery BIO PolyMA-WAX Columns
Column:   Discovery BIO PolyMA-WAX, 5cm x 4.6mm, 5µm (Cat. No. 59602-U)
Mobile Phase:   (A) 20mM Tris-HCl, pH 8.0;
(B) 20mM Tris-HCl, 0.5M NaCl, pH 8.0
Flow:   0.5mL/min
Temp.:   25°C
Detection:   UV, 280nm
Sample:   Myoglobin (5µg), conalbumin (5µg), trypsin inhibitor (10µg)
Gradient Profile:  
time (min.) %A %B
0 95 5
15 0 100

 


Ordering Information
Discovery BIO PolyMA-WAX HPLC Column
Particle Size (µm) ID (mm) Length (cm) Cat. No.
5 4.6 5 59602-U
 
Column Protection: We recommend filtering samples and using an in-line filter to protect Discovery BIO PolyMA-WAX columns.
Description Cat. No.
In-line filter, 0.5 micron frits Z227323

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TRADEMARKS
Discovery is a trademark of Sigma-Aldrich Co.