Papain digestion of humanized monoclonal IgG was analyzed using PROTEIN LW-803, a column for aqueous SEC (GFC). Fragments such as Fc and Fab constituting IgG, and transient decomposition intermediates are assumed to emerged throughout the degradation process of IgG with Papain. LW-803 is suitable for the separation of IgG’s components, fragments and intermediates produced by decomposition. Furthermore, LW-803 improved the separation performance for the molecular weight range around IgG. So LW-803 is effective not only for the analysis of aggregates (oligomers), which are polymer side impurities in antibody production, but also for analysis of low molecular components and monitoring of fragmentation.

（Procedure of papain digestion）
(1) Dissolve 3 mg of humanized monoclonal IgG in 500 µL of the eluent. (6 mg/mL conc.)
(2) Dissolve 1 mg of papain in 500 µL of the eluent. (1 mg/mL conc.)
(3) Pass (1) and (2) through 0.2 µm membrane filter.
(4) Mix each solution in equal amounts.
(5) Keep temperature at 25 °C.
(6) Takes a sample with time and analyze it by HPLC.


Sample : 10 μL
Humanized monoclonal IgG
1. Aggregates of IgG
2. Dimer of IgG
3. Monomer of IgG
4, 5, 6. Fragments of IgG
from papain digestion
7. Citric acid
8. Papain

Column       : Shodex PROTEIN LW-803 (8.0 mm I.D. x 300 mm)
Eluent       : 0.1 M Sodium phosphate buffer (pH7.0) + 0.3 M NaCl
Flow rate    : 1.0 mL/min
Detector     : UV (280 nm)
Column temp. : 25 °C