STATIC PERFORMANCE OF MABXPURE

APPLICATION NOTE # 1

DAISEP MabXpureTM, a disposable bead-based multimodal (SEC-AEX)
flowthrough technology offering high depletion performances for Host
Cell Proteins and DNA during clarification/harvest stages and sample
preparation steps for mAb product solutions.

INTRODUCTION
Today, biopharmaceutical manufacturing has shown
major improvements in upstream efficiency, withincreasing titers (> 10 g/L) generally associated withhigh cell densities (> 60 x 10e6 cells/mL). Such increasein productivity is creating high levels of process-related
impurities like greater HCP levels (> 800.000 ng/mL),
host cell DNA (> 1000 ng/mL) content, HMW, fragments
and/or lipids. This increase in bioburden and impurity
levels shifts the production bottleneck to purification(clarification, chromatography and filtration). In this
scenario, centrifugation and depth filtration techniques
are reaching their limits, unable to provide a sustainablepurification platform in terms of impurity reduction
and/or bioburden depletion. DAISEP MabXpure has

been developed to target these scenarios to ensuresignificant bioburden depletion with sustainableefficiencies of clarification. DAISEP MabXpure utilizes acombination of anion exchange (AEX) and size
exclusion (SEC) mechanisms to trap the impurities
whilst retaining mAbs in the flowthrough (Figure 1.). The
bead design has been adapted to enable single-use
flowthrough harvest operations using MabXpure as afilter aid in combination with depth filters (Figure 2.).
DAISEP MabXpure can be mixed directly with the mAb
feedstock to implement the bioburden depletion
process. DAISEP MabXpure’s high selectivity forimpurities allows high HCP and DNA depletion (> 1 LRV)
with high mAb recoveries (> 95%).

SEC

DAISEP MabXpure
mAbHCPs

Centrifuge

Alluvial Depth
Filtration

FeedHCDNA

AEXDirect
Filtration

Smallmolecules
nucleic acids

Figure 1. Multimodal depletion mechanisms ofDAISEP MabXpure

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please contact your local office.

Filter aid(MabXpure, DE)
Cakeformation

Figure 2. Cake / Alluvial filtration scheme

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STATIC PERFORMANCE OF MABXPURE

APPLICATION NOTE # 1

MATERIALS AND METHODS:
LOAD MATERIALBUFFER

DEVICE

CHO-K1 clarified feed, spiked
with mAb 2 g/L, HCP > 200 μg/mL,
DNA > 1,000 ng/mL

Tris HCl 50 mM
pH 7.4

MabXpure,
DAISEP Spincentrifuge tubes

Depletion is performed by enabling product contact withthe resin in different mix ratios i.e different ratios ofDAISEP MabXpure with the CHO-K1 feed during 5
minutes contact time, and filtering off the resin withDAISEP Spin centrifuge tubes. Following this centrifugal

filtration process, quantification of HCP is performed
using the CHO-specific ELISA kits from Cygnus, DNA
is quantified with Picogreen reagent and mAb titer ismeasured by HPLC-SEC. Additional contact times wereutilized for 1:50 mix ratio.

RESULTS
In conventional clarification processes, where classic filter aids are used (silica or diatomaceous earth DE), either
bioburden depletion is effective or there is an effective mAb recovery obtained (Figure 3.). There is always a trade-off on
recovery or bioburden depletion depending on which one takes precedence. However, with DAISEP MabXpure there is
combined efficacy in terms of bioburden removal (> 1 LRV) and antibody recovery (> 80%). These performances areguaranteed and observed even when DAISEP MabXpure is diluted up to 1:20 mix ratio, representing ideal conditions forsample preparation and harvest process operations (Figure 3.). If necessary, the depletion performances can beincreased with higher contact times (Figure 4.) inducing lower residual HCP and by reaching 2 LRV of DNA.

100%

100100100

1009790%
70%

40%

10%
Feed40%37

97

1:5 Silica 1:5 DEHCPs

9950%

2620%

9960%

3530%

10070%

6050%

100100100

80%836960%

100%
90%

8880%

9829125

1:5

1:10DNAmAb

91:20

Figure 3. Residual HCP, DNA and mAb recoverywith different mix ratios of DAISEP MabXpure

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please contact your local office.

3730%
232320%10%

1:50

FeedHCPs

5’2724
3

130’

120’

DNAmAbFigure 4. Impact of contact time on residual HCP,DNA and mAb recovery (1:50 mix ratio)

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STATIC PERFORMANCE OF MABXPURE

APPLICATION NOTE # 1

DISCUSSION:The use of MabXpure in static mode fits naturally in aprocess where depth filters are used since the resin iseliminated by cake filtration or alluvial depth filtration. As aconsequence, the number of depth filter sheets is adapted
to the amount of filter aid to be removed.

clearance < 10 NTU starting with feeds > 4000 NTU. Ideally,
DAISEP MabXpure’s conditions of use are optimal with highmix ratios and short contact times for sample preparation, or
lower mix ratios and higher contact times for in-processapplications. The flexibility of utilising the resin in the process,in this manner, enhances its single-use and flowthroughquality attributes enabling the ease of implementation ofMabXpure in the process.

DAISEP MabXpure can also be combined with diatomaceous
earth with unclarified feedstocks achieving good turbidity

CONCLUSIONDAISEP MabXpure has a very high capability for
depleting host cell proteins and DNA, withextremely high mAb recovery.

Depending on the process conditions, the capacityof DAISEP MabXpure helps to remove up to 1 LRVof HCP and 2 LRV of DNA in one static mode step.

The unique flexibility of DAISEP MabXpure allows
the user to choose the mode of action with anoptimal mix ratio and contact time to make theresin an ideal filter aid for clarification/harvest
steps by cake or alluvial filtration.

DAISEP MabXpure can be implemented from lab
scale to process scale with DAISEP Spin columns
to depth filter sheets respectively.

XS

BATCH
INCUBATIONCHAIMBERRESIN
YOUR SAMPLE

PURIFIED PROTEIN

To place an order or receive technical support,
please contact your local office.

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STATIC PERFORMANCE OF MABXPURE

APPLICATION NOTE # 1

DAISEP MABXPURE RANGEMABXPURE BULK
MabXpure bulk 50 mLDMXBK0050

MabXpure bulk 500 mLDMXBK0500

MabXpure bulk 1000 mLDMXBK1000

MabXpure bulk 10L

DMXBK1050 mL

500 mLMABXPURE FT
MabXpure FT 1 mL (5 units/pack)

DMXFT0001

MabXpure FT 5 mL (5 units/pack)

DMXFT0005

MabXpure FT 50 mL (1 unit)

DMXFT0050

MabXpure FT 500 mL (1 unit)

DMXFT0500

MabXpure FT 1.5 L (1 unit)

DMXFT1500

1 mL

5 mL

MABXPURE KIT
MabXpure Kit

DMXKT0001
MABXPURE KITDAISEP SPIN XS & XL
DAISEP Spin XS (40 units/pack)

DSPXS0040

DAISEP Spin XL (8 units/pack)

DSPXL0008XSXLDAICEL BIOSEPARATIONS - LOCATIONSNORTH/LATIN AMERICACHINAEUROPEINDIA

Chiral Technologies, Inc.
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