Bio LC Columns
Shim-pack Scepter C4
Design for biomolecules separations Shim-pack Scepter C4 is a wide-bore C4 column that uses an organic silica hybrid base which is suitable for separation of proteins such as monoclonal antibodies. The column can be used for analysis of proteins with a molecular weight of up to approximately 150,000 and is suitable for analysis of antibodies as it is very stable even under acidic or high- temperature conditions because an organic silica hybrid base
Bonded Phase
Butyl Groups
Particle Size
1.9μm, 3μm, 5μm
Pore Size
30 nm
End-capping
Yes
pH Range USP Code
1 to 10
L26
is used. It is also effective for high-sensitivity analysis on LC/MS as it provides good peak shapes even when using a formic acid mobile phase with low ion paring effect. In addition, it is effective for separation of highly-hydrophobic proteins, nucleotides and small molecules that are hard to separate using a regular C18 column.
Analysis Examples
Analysis of Proteins
Analysis of Monoclonal Antibodies
Peaks 1. Ribonuclease A
(bovine pancreas)
2. Cytochrome C (horse heart) 3. Insulin (bovine pancreas) 4. BSA 5. α -Chymotrypsinogen A (bovine pancreas)
NIST mAb Bevacizumab Adalimumab Rituximab Trastuzumab
Conditions
Conditions
Column
Column
: Shim-pack Scepter C4 (1.9 μm, 50 x 2.1 mm I.D.) (P/N: 227-31175-03)
: Shim-pack Scepter C4 (5 μm, 150 x 3.0 mm I.D.) (P/N: 227-31181-04)
Mobile Phase : A) Water/TFA (100/0.1)
Mobile Phase : A) Water/TFA (100/0.1)
B) Acetonitrile/TFA (100/0.1) 25-45%B (0-10 min)
B) Acetonitrile/TFA (100/0.1) 20-60%B (0-27 min), 90%B (27-35 min), 20%B (35-50 min)
Flow Rate Col. Temp. Detection
: 0.4 mL/min
Flow Rate Col. Temp.
: 80 ºC
: 0.4 mL/min
: UV at 280 nm
: 70 ºC
Injection Vol. : 2 µL Sample Conc. : 0.2 mg/mL
Detection : UV at 220 nm Injection Vol. : 10 µL (0.25 – 0.50 mg/mL) Column
Pg A-22
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