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| Discovery BIO PolyMA-SCX |
| Polymethacrylate polymer-based strong cation-exchange
HPLC column |
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Discovery BIO PolyMA-SCX polymer-based cation-exchange
particles have discriminating hydrophilic surface chemistry making them
ideally suited for separating proteins, peptides, and other biotechnology-derived
products.
Choose Discovery BIO PolyMA-SCX for cation-exchange separations. Generally run at
a pH less than the protein's pI (isoelectric point) - usually below pH 7. |
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| Particle: |
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Polymethacrylate with hydrophilic coating |
| Functional Group: |
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Sulfopropyl (SP) |
| Counterion (as supplied): |
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Na+ |
| 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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1 to 13 |
| Temperature Range: |
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4°C to 50°C |
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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 CATION-EXCHANGE SEPARATIONS |
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| Discovery BIO PolyMA-SCX typically gives better resolution
than competitive polymeric ion-exchange materials. |
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| Comparison of Efficiency: Cytochrome
c Variants |
| Columns: |
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Discovery BIO PolyMA-SCX, 5cm x 4.6mm,
5µm (Cat. No. 59601-U) and Competitive polymeric-SCX columns
of similar dimensions |
| Mobile Phase: |
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(A) 50mM MOPS/KOH, pH 7.0;
(B) 50mM MOPS/KOH, 0.5M KCl, pH 7.0 |
| 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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10µg each variant |
| Gradient Profile: |
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0.6% B per minute. See Figure for details. |
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| 1. |
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Cow cytochrome c |
| 2. |
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Horse cytochrome c |
| 3. |
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Rabbit cytochrome c |
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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-SCX columns. |
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| Ion-exchange Capacity |
| Column |
Protein |
mg per mL of column volume |
mg per column |
| Discovery BIO PolyMA-SCX |
Lysozyme |
40mg |
30mg |
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| Loading studies were run at 240cm/hr (0.67mL/min) using
0-1M NaCl salt gradients in 20mM sodium phosphate, pH 7. Protein concentration
was 2mg/mL in starting mobile phase. |
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| Protein (Fibrinogen) Recovery |
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Cytochrome C Recovery |
| Column |
1st Injection |
2nd Injection |
5th Injection |
| Discovery BIO PolyMA-SCX (5µm, 5cm
x 4.6mm) |
99.5% |
98.6% |
100.2% |
| Competitive polymethacrylic SP column
(10µm, 7.5cm x 7.5mm) |
59.0% |
76.1% |
90.6% |
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| Mobile phase: Gradient of 0 - 1M NaCl in 20mM citrate
(pH 4) over 30 minutes. Flow Rate: 1mL/min. Ambient temperature. Sample: 40µg cytochrome c 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 will be reproducible injection after injection because of
exceptional column stability. In the figure below, Discovery BIO PolyMA-SCX
columns are shown to give stable retention of three proteins after 500 injections,
with no sign of deterioration. |
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| Stability of Discovery BIO PolyMA-SCX
Columns |
| Column: |
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Discovery BIO PolyMA-SCX, 5cm x 4.6mm,
5µm (Cat. No. 59601-U) |
| Mobile Phase: |
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(A) 20mM sodium phosphate, pH 7.0
(B) 20mM sodium phosphate, 0.5M NaCl, pH 7.0 |
| 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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Ribonuclease A (10µg), a-chymotrypsinogen
(5µg), cytochrome c (5µg) |
| Gradient
Profile: |
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| time (min.) |
%A |
%B |
| 0 |
95 |
5 |
| 15 |
0 |
100 |
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