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Applications

IgG_Dog (Native) Jupiter 300 C4

Application Detail (App ID: 14908)

Part No:00F-4167-B0
Dimensions:150 x 2 mm ID
Elution Type:Gradient
Elution A:0.1%TFA/95% Water/ 5% Acetonitrile
Elution B:0.085% TFA/75% Acetonitrile/ 20% IPA/5% Water
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 80 20
2 20 10 90

Flow Rate:0.25 mL/min
Col. Temp: ambient
Detection: UV-Vis Abs.-Diode Array (PDA) @ nm (22°C)
Note:These applications demonstrate the utility of using low amounts of isopropanol in the organic mobile phase to improve peak shape of hydrophobic proteins. App ID#14918 shows the elution of Ig-G on a Jupiter 300 C4 column where Acetonitrile is used as the organic mobile phase. App ID#14908 shows the same Ig-G run on the same column using an organic mobile phase that contains 20% isopropanol. Note the dramatic improvement in both recovery and peak shape for the Ig-G peak. The addition of isopropanol to the organic mobile phase improves the efficiency as well as the peak shape for large hydrophobic proteins due much in part to the increased hydrophobicity of the organic mobile phase upon addition of isopropanol. Especially when performing separations of Ig-G and other immunoglobin one should start with the use of a wide pore C4 column and look at using significant amounts of IPA as part of the organic mobile phase.

Analyte Details

Analyte Detail not available

1
IgG
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