Lifetime study for JSR Amsphere A3 – protein A resin
Posted on January 27,2017
Many factors are required to assess and implement a protein A resin into a mAb purification process. Dynamic binding capacity, purity, operating flow rate and caustic compatibility are a few among many evaluation parameters. Resin lifetime is a key parameter that needs to be assessed as it is directly related to both resin volume and process costs. The resin should maintain performance parameters over the intended number of process cycles. The following describes the effects of process cycling and caustic CIP on the purification performance of Amsphere A3 over 100 cycles.
Amsphere A3 is a new protein A resin designed with a surface modified base bead and alkali-resistant optimized ligand.
Protein A ligand
• High DBC via controlled conformation and orientation
• High alkaline stability from protein engineering Surface modification
• Low HCP levels by surface hydrophilization Base bead formulation
• High DBC at high flow rate
• Excellent pressure and flow properties via rigid crosslinking
Materials and Methods
• Biosimilar mAb, 1.1 g/l titer, pI 8.45, from CHO cell line
• Initial HCP level 350,000 ppm
• Clarified by centrifugation and 0.22 micro filtration
• HCP determination by Cygnus Kit (F550)
• DNA determination by Picogreen® assay (Invitrogen)
• Leached protein A determination by Cygnus Kit (F740)
TABLE 1: CIP CYCLE TEST CONDITIONS
FIGURE 1: TEST CONDITIONS OF CHROMATOGRAPHY PROCESS
1. DYNAMIC BINDING C
2. mAb RECOVERY
3. HCP REDUCTION
4. DNA REDUCTION
5. LEACHED PROTEIN A INTO TARGET POOL
6. ELUTION POOL
7/8. CHROMATOGRAMS COMPARISON
9. ELUTION PROFILE
The CIP test results indicate stable performance over 100 operating cycles, including NaOH exposure of 0.1 M NaOH every cycle and 0.5 M NaOH every tenth cycle. The mAb recovery, HCP reduction, DNA reduction and elution pool pH are consistent across the 100 cycles. The chromatograms from the first to the 101st cycle also show nearly identical response both in terms of elution profile and column volume.
This data set indicates stable resin performance can be expected after multiple cycles.
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