Mixed Mode Chromatography & KBI's mAB Purification Platform

1. 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
1
DS
Process
Platform
Cell line
diversity
Media/feed
type
diversity
HCP level
variability
Cell density
variability
HMW level
variability

2. 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
2

3. Mixed-mode cation exchange/hydrophobic
interaction resins
3
NuviaTM cPrimeTM
Bio-Rad Laboratories
MX-Trp-650M
Tosoh
EshmunoTM HCX
EMD Millipore
CaptoTM MMC
GE Healthcare
TM

4. High Throughput Screening: Plate Experimental Design
4/27/2016
4
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

5. 5
• 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

6. 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
6