Biosolutions and Columns for Biologic Characterization
Separation of Protein Standards on the ZORBAX GF-250 SEC Column
5
Column:
ZORBAX GF-250 884973-901 9.4 x 250 mm, 4 μm
Mobile Phase: Flow Rate: Temperature: Detector: Sample:
200 mM Sodium Phosphate, pH 7.0 2 mL/min Ambient 254 nm BioRad Gel Filtration Standards for Size Exclusion 1. Thyroglobulin 670,000 Da 2. Bovine Gamma Globulin 158,000 Da 3. Chicken Ovalbumin 44,000 Da 4. Equine Myoglobin 17,000 Da 5. Vitamin B-12 1,350 Da
4
1
2
The protein standards separated here are a commonly selected set of standards. The ZORBAX GF-250 column shows excellent resolution for this sample. Additional resolution of the thyroglobulin can be obtained by adding the GF-450 column in series.
0
1
2
3
4 Time (min)
3
5
6
High-Speed Size Exclusion Separations 2
Column:
ZORBAX GF-450 884973-902 9.4 x 250 mm, 6 μm
Column:
ZORBAX GF-450 884973-902 9.4 x 250 mm, 6 μm
Mobile Phase:
PBS (phosphate buffered saline), pH 7.4 3 mL/min Ambient 220 nm BSA and BSA Dimers
Mobile Phase:
200 mM Na Phosphate Monobasic pH 7.0/0.1% Azide 2 mL/min 225 nm 10 μg in 50 mM Sodium Phosphate pH 7.0
Flow Rate: Detector: Sample:
BSA and BSA Dimers (duplicate injections)
Antibody Separation 2
2
2
1. IgG Dimer and Aggregate 2. Mouse IgG 1K (MOPC 31C) 3. Salts: Tris, Na2HPO4, NaCl
1. BSA dimer 2. BSA monomer 3. Azide 1 Inj. #1
0
1002
1
3
LC AND LC/MS
3
3
Inj. #2
5 Time (min)
8 LCGF001
High-Speed Size Exclusion Separations 1
Flow Rate: Temperature: Detector: Sample:
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1 10
www.agilent.com/chem/lccolumns
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4 5 6 7 Retention Time (min)
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10 LCGF002