IgG_Dog (Native) Jupiter 300 C4

LC Conditions

Column

Jupiter 5 µm C4 300 Å, LC Column 150 x 2 mm, Ea

Brand

Jupiter

Part No

00F-4167-B0

Phase Name

C4

Elution Type

Gradient

Mobile Phase

A: 0.1%TFA/95% Water/ 5% Acetonitrile

B: 0.085% TFA/75% Acetonitrile/ 20% IPA/5% Water

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

80

20

2

20

10

90

Flow Rate

0.25 mL/min

Temperature

22°C

Detection

UV-Vis Abs.-Diode Array (PDA) @ (22°C)

Detection Info

Sample Notes

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.