Monoclonal Antibody Purification and Quantification Using MPC and CHT Media
MAb-S Purification Profile
Fig. 1A. The chromatographic performance for purifying the MAb-S sample with MPC (
) and CHT Type I, 40 µm () shows separation of the monomer from higher molecular weight impurities.
SEC Profile of Pooled Monomer Fractions
Fig. 1B. The elution pools collected on the MAb-S antibody sample purified on MPC and CHT Type I, 40 µm (in Figure 1A above) were analyzed for purity by size exclusion chromatography. The overlaid chromatograms of the elution pools show only one major peak and no evidence of dimers or aggregates in the MPC ( ) or CHT () separations.
Data Quantification
| |
IgG concentration, mg/ml |
Protein A, ppm |
HCP, ppm |
DNA, ppm |
Dimer/aggregate, % |
| MAb-S |
|
|
|
|
|
| Protein A eluate |
2.90 |
33 |
24 |
21 |
21.62 |
| CHT pool |
0.40 |
<0.6 |
<2 |
2.62 |
<0.03 |
| MPC pool |
0.46 |
<0.5 |
<2 |
2.83 |
<0.03 |
Table 1. All pooled fractions and starting material from MAb-S purified on MPC and CHT Type I, 40 µm (Figure 1B above) were analyzed for host cell protein using a Cygnus Technologies HCP ELISA kit, for dsDNA using a PicoGreen dsDNA quantitation kit from Molecular Probes, Inc., for Protein A leachables using a Cygnus Technologies Protein A ELISA kit, and for aggregate clearance by size exclusion chromatography on a Bio-Sil® SEC 250 column.
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