PEGylated proteins on Jupiter® 300 C18 3µm and 5µm, and C4 5µm columns

LC Conditions

Column

Jupiter 3 µm C18 300 Å, LC Column 150 x 4.6 mm, Ea

Brand

Jupiter

Part No

00F-4263-E0

Phase Name

C18

Elution Type

Gradient

Mobile Phase

A: 0.1% TFA, 2% ACN in Water

B: 90% ACN, 0.085% TFA in Water

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

80

20

2

25

35

65

Flow Rate

1.00 mL/min

Temperature

45°C

Detection

UV-Vis Abs.-Variable Wave.(UV) @ (25°C)

Detection Info

Sample Notes

Compare reverse phase separations of different PEGylated and intact proteins selected at different reaction time-points on three different columns. (A) PEGylated alpha-Chymotrypsinogen A with PEG1 Amine 20kDa Methoxy PEG NHS-ester, 0 min fraction; (B) PEGylated alpha-Chymotrypsinogen A with PEG3 N-Terminus 20kDa Methoxy PEG aldehyde, 120 min fraction; (C) PEGylated beta-Lactoglobulin A with PEG4 N-Terminus 40kDa Y-shape Methoxy PEG aldehyde, overnight fraction. Well-noticed, good resolution was achieved on Jupiter 3µm C18 small particles versus larger Jupiter 5µm C18 particles or Jupiter C4 bonded media.