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Comparison of SEC Separation of Standard Proteins


Standard proteins were analyzed with PROTEIN LW-803 which is a column for Aqueous SEC (GFC). PROTEIN LW-803 is suitable for analyzing proteins with a molecular weight of several hundreds of thousands. Furthermore, this column is especially suitable for separating γ-Globulin with the molecular weight of 160,000 and vicinity. The samples are better separated with LW-803 than a conventional column (KW-803) 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
1. Thyroglobulin
(MW : 670,000) 7mg/mL
2. γ-Globulin
(MW : 160,000) 6mg/mL
3. Ovalbumin
(MW : 44,300) 4.8mg/mL
4. Ribonuclease A
(MW : 13,700) 7mg/mL
5. Uridine
(MW : 244) 0.1mg/mL

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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