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| Discovery BIO PolyMA-WAX |
| Polymethacrylate polymer-based weak anion-exchange HPLC
column |
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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.
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| Particle: |
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Polymethacrylate with hydrophilic coating |
| Functional Group: |
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Diethylaminoethyl (DEAE) |
| Counterion (as supplied): |
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Cl- |
| Particle Shape: |
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Spherical, monodispersed |
| Particle Size: |
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5µm |
| Pore Size: |
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1000Å |
| Coverage: |
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0.3meq/g |
| pH Range: |
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2 to 10* |
| Temperature Range: |
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4°C to 50°C |
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| * PolyMA-WAX is a weak anion-exchange material. It can be
used at high pH values but with reduced charge. |
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| Solutions for Protein and Peptide Separation Challenges |
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Excellent separations of protein isoforms |
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High resolution at low sample load |
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Quantitative recovery - a hydrophilic
surface eliminates protein adsorption |
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High efficiency |
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Wide pH range |
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| EFFICIENT ANION-EXCHANGE SEPARATIONS |
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| Discovery BIO PolyMA-WAX typically has higher efficiency
than competitive polymeric ion-exchange materials. |
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| Comparison of Efficiency: Hemoglobin
Variants |
| Columns: |
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Discovery BIO PolyMA-WAX, 5cm x 4.6mm,
5µm (Cat. No. 59602-U) and Competitive polymeric weak anion
exchange columns |
| Mobile Phase: |
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(A) 10mM Tris/HOAc, pH 8.0;
(B) 10mM Tris/HOAc, 0.25M KCl, pH 8.0 |
| Flow: |
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3.01cm/min (flow rates appear in Figure) |
| Temp.: |
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35°C |
| Detection: |
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UV, 280nm |
| Sample: |
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50µg each variant |
| Gradient Profile: |
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1.6% B per minute. See Figure for details. |
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| 1. |
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Hemoglobin A2 |
| 2. |
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Hemoglobin S |
| 3. |
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Hemoglobin A0 |
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| BALANCE OF ION-EXCHANGE CAPACITY AND PROTEIN RECOVERY |
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| 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. |
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| Ion-exchange Capacity |
| Column |
Protein |
mg per mL of column volume |
mg per column |
| Discovery BIO PolyMA-WAX |
BSA |
50mg |
40mg |
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| 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. |
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| Protein (Fibrinogen)
Recovery |
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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% |
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| 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. |
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| COLUMN STABILITY AND LIFETIME |
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| 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. |
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| Stability of Discovery BIO PolyMA-WAX
Columns |
| Column: |
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Discovery BIO PolyMA-WAX, 5cm x 4.6mm,
5µm (Cat. No. 59602-U) |
| Mobile Phase: |
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(A) 20mM Tris-HCl, pH 8.0;
(B) 20mM Tris-HCl, 0.5M NaCl, pH 8.0 |
| Flow: |
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0.5mL/min |
| Temp.: |
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25°C |
| Detection: |
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UV, 280nm |
| Sample: |
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Myoglobin (5µg), conalbumin (5µg),
trypsin inhibitor (10µg) |
| Gradient
Profile: |
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| time (min.) |
%A |
%B |
| 0 |
95 |
5 |
| 15 |
0 |
100 |
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