a comparison of multimodal chromatography resin: case study
TRANSCRIPT
Mixed Mode Chromatography and KBI’s mAB Purification Process Platform• Mixed Mode resins are ideal for KBI’s mAb platform process since by the nature of
our business there is a significant diversity in mammalian cell clones/cell culture harvest and mAbs
• Mixed mode resins can provide added robustness to a mAb platform to reduce both HMW and HCP impurities
• Given the complexity of mixed mode resin interactions, optimizing a mixed mode chromatography step can be time consuming and potentially material limiting
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DS ProcessPlatform
Cell line diversity
Media/feed type
diversity
HCP level variability
Cell density variability
HMW level variability
Objectives• Determine if there is at least one mixed mode cation
exchange/hydrophobic interaction polishing step resin that performs well for multiple mAbs• Ideal candidates for a mAb process platform
• Present a high throughput, low material requirement method for identifying optimal operating ranges for multiple mixed mode resins in a single set of experiments• Show how the method can be use to identify optimal
chromatography conditions
• Show the relationship between the high throughput screening data and chromatography performance for different mAbs
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Mixed-mode cation exchange/hydrophobic interaction resins
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NuviaTM cPrimeTM
Bio-Rad Laboratories
MX-Trp-650MTosoh
EshmunoTM HCXEMD Millipore
CaptoTM MMCGE Healthcare
TM
High Throughput Screening: Plate Experimental Design
05/03/2023
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Variable Conditions Evaluated
Load pH 4, 5, 6, 7, 8
Elution pH 4, 5, 6, 7, 8
Elution [NaCl] (M) 0.20, 0.65, 1.10, 1.55, 2.00
• Three mAbs evaluated under a series of conditions• One plate executed per load pH
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• The low yield process condition zone size varies significantly with respect to resin, mAbs and process conditions and are easily identifiable
• In general, Capto MMC has slightly larger low yield zones for the mAbs and test conditions within our typical operating pH range .
• In general, each resin has a characteristic surface response map profile
Findings regarding the relationship between HT filter plate and chromatography data• High throughput filter plate data can be used to quickly
limit the focus of a chromatography evaluation using minimal amounts of the target protein• pH and NaCl conditions not favorable for high yields
are easily identifiable• There is a correlation between yield variability for each
condition (load pH and elution pH) and monomer selectivity• more variability leads to better selectivity
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