BioHPLC Column Selection Guide Cover - Agilent Technologies
BioHPLC Column Selection Guide Cover - Agilent Technologies
BioHPLC Column Selection Guide Cover - Agilent Technologies
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Comparison of <strong>Agilent</strong> Bio SEC-3<br />
and competitor column in the analysis<br />
of a monoclonal antibody<br />
<strong>Column</strong>: Bio SEC-3, 300Å<br />
5190-2511<br />
7.8 x 300 mm, 3 µm<br />
<strong>Column</strong>: Competitor 7.8 x 300 mm<br />
Mobile Phase: 150 mM sodium phosphate<br />
+ 100 mM Na sulfate (with salt)<br />
150 mM sodium phosphate (without salt)<br />
Flow Rate: 1.0 mL/min<br />
Detector: 220 nm<br />
Sample: MAb (2 mg/mL)<br />
mAU<br />
40<br />
30<br />
20<br />
10<br />
0<br />
With salt<br />
Without salt<br />
<strong>Agilent</strong> Bio SEC-3<br />
Monomer<br />
Dimer<br />
size exclusion Chromatography (seC)<br />
8 minutes<br />
0 2 4 6 8 10 min<br />
mAU<br />
40<br />
The <strong>Agilent</strong> Bio SEC-3 column reveals the presence of MAb fragments missed by the competitor column.<br />
30<br />
20<br />
10<br />
0<br />
Competitor column<br />
Monoclonal Antibody Monomer and Dimer Analysis<br />
using <strong>Agilent</strong> Bio SEC-3 and a Competitor <strong>Column</strong><br />
Eluent <strong>Column</strong><br />
Resolution Ratio<br />
Monomer:Dimer<br />
With salt<br />
Without salt<br />
Monomer<br />
Dimer<br />
8 minutes<br />
0 2 4 6 8 10 min<br />
Monomer<br />
Efficiency<br />
Mono_Dimer1<br />
Percentage<br />
Dimer<br />
With salt <strong>Agilent</strong> 2.04 7,518 0.59<br />
With salt Competitor 1.88 3,967 0.59<br />
Without salt <strong>Agilent</strong> 2.08 7,942 0.60<br />
Without salt Competitor 1.92 4,164 0.57<br />
UHPLC/HPLC<br />
9
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