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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