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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Monoclonal IgG1 chains:<br />
Separation on Poroshell 300SB-C8<br />
<strong>Column</strong>: Poroshell 300SB-C8<br />
660750-906<br />
2.1 x 75 mm, 5 µm<br />
Mobile Phase: A: 90% water:<br />
10% ACN + 3 mL/L of MW 300 PEG<br />
B: 10% water:<br />
90% ACN + 3 mL/L of MW 300 PEG<br />
Flow Rate: 1.0 mL/min<br />
Gradient: 0 min 25% B<br />
10 min 40% B<br />
10.1 min 25% B<br />
12 min 25% B<br />
Temperature: 70 °C<br />
Sample: Monoclonal IgG1<br />
Courtesy of:<br />
Novartis Pharma,<br />
Biotechnology, Basel<br />
Dr. Kurt Forrer<br />
Patrik Roethlisberger<br />
mAU<br />
120<br />
80<br />
40<br />
0<br />
0<br />
mAU<br />
2 4 6 8 10<br />
120<br />
Glycosylated?<br />
Non-Glycosylated?<br />
80<br />
40<br />
0<br />
0<br />
mAU<br />
2 4 6 8 10<br />
120<br />
80<br />
40<br />
0<br />
0 2 4 6 8 10<br />
Tips & Tools<br />
Light Chains<br />
Heavy Chains<br />
Reversed-Phase HPLC<br />
IgG treated with:<br />
DTT<br />
IgG treated with:<br />
DTT<br />
Peptide-N-Glycosidase F<br />
IgG treated with:<br />
DTT<br />
Peptide-N-Glycosidase F<br />
Carboxypeptidase-B<br />
agilent offers an extensive selection of certified chromatography<br />
sample vials including polypropylene and deactivated and<br />
siliconized glass. For more information see 5990-90322en<br />
www.agilent.com/chem/library<br />
UHPLC/HPLC<br />
m<br />
m<br />
m<br />
IgG_Porosh<br />
33
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