Separation of Mouse Ascites Fluid Containing Monoclonal Antibodies and purified Monoclonal Antibodies
Columns: |
TSKgel Q-STAT, 7 µm,
4.6 mm ID × 10 cm |
Mobile phase: |
A: 20 mmol/L Tris-HCl buffer, pH 8.5
B: 0.5 mol/L NaCl in 20 mmol/L Tris-HCl buffer, pH 8.5 |
Gradient: |
A to B linear gradient (10 min) |
Flow rate: |
1.0 mL/min |
Detection: |
UV @ 280 nm |
Temperature: |
25 °C
|
Injection vol.: |
10 µL
|
Sample: |
Top: 1/10 dilution of mouse ascites containing mAb
Bottom: purified mouse mAb
Sample diluted 10-fold with eluent A |
Separation of Proteins Within 1 Minute
Column: |
A. TSKgel Q-STAT, 10 µm,
3 mm ID × 3.5 cm
B. ProSwift WAX-1S Monolith
4 mm ID × 5 cm
|
Mobile phase: |
A: 20 mmol/L Tris-HCl, pH 8.5
B: 0.5 mol/L NaCl in buffer A |
Gradient: |
0 min (0% B), 1 min (100% B) |
Flow rate: |
2.0 mL/min |
Detection: |
UV @ 280 nm |
Samples: |
1. conalbumin
2. ovalbumin
3. trypsin inhibitor |
|
|
Analysis of IgG
Column: |
A: TSKgel Q-STAT, 7 µm,
4.6 mm ID × 10 cm
B: ProPac® WAX-10, 10 µm, 4 mm ID × 25 cm |
Mobile phase: |
A: 20 mmol/L Tris-HCl, pH 8.5
B: 0.5 mol/L NaCl in buffer A |
Gradient: |
0 min (0% B) 10 min (100% B) |
Flow rate: |
1.0 mL/min |
Detection: |
UV @ 280 nm |
Sample: |
pepsin digested mAb |
|
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