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Comparison of SEC Separation of Polyclonal Antibody


Polyclonal antibody (IgG form human serum) was analyzed with PROTEIN LW-803 which is a column for Aqueous SEC (GFC). The LW-803 column is similar to KW-803, a conventional column. γ-Globulin with the molecular weight of 160,000 and vicinity are better separated with LW-803 than the conventional column or other manufacturer’s columns. Improvement of separation performance for this molecular weight range is effective for separation of monomer and dimer of IgG which is the mainstream of antibody drugs.

Sample : 5μL
IgG from human serum 10mg/mL
1. Aggregates
2. Dimer
3. Monomer

Columns      : Shodex PROTEIN LW-803 (8.0mmI.D. x 300mm)
               Shodex PROTEIN KW-803 (8.0mmI.D. x 300mm)
               Silica-based SEC column from other manufacturer (7.8mmI.D. x 300mm)
Eluent       : 50mM Sodium phosphate buffer (pH7.0) + 0.3M NaCl
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

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