DYNAMIC PERFORMANCE OF MABXPURE FTAPPLICATION NOTE #2
DAISEP MabXpure™, a single-use multimodal (SEC-AEX)
flowthrough (FT) technology application offering a high performance solution for the depletion and reduction of Host Cell Proteins(HCPs) and DNA following clarification / harvest, during purification,
polishing and sample preparation of MAbs and large molecules.

INTRODUCTION
Today, biopharmaceutical manufacturing has shownconsiderable improvements in upstream processing
efficiencies, enabling high process titers in excess of 10 g/L
associated with high cell densities (>60 x 10e6 cells/mL). This
increase in productivity also creates high levels of processrelated impurities like higher HCP levels (>800.000 ng/mL),
host cell DNA (>1000 ng/mL) content, HMW, fragments and/or lipids. The huge increase in bioburden and impurity levels
shifts the production bottleneck to purification (clarification,chromatography and filtration).
In the current scenario, primary separation techniques likecentrifugation, depth filtration combined with secondary
purification operations are reaching their limits. Conventional
bioseparation technologies are unable to provide an efficient,consistent and sustainable purification platform in terms of
impurity reduction and/or bioburden depletion.
Recently, there has been considerable focus on thedevelopment of new single-use flowthrough technologies
since they can provide a unique flexibility of being easy toFigure 1. Multimodal depletion mechanisms of DAISEP MabXpure

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

implement with excellent process performance and achieving
high process recoveries. DAISEP MabXpure is a prefilled disposable device which is developed based on theaforementioned performance criteria to ensure significant
bioburden depletion using a combination of anion exchange(AEX) and size exclusion (SEC) mechanisms.
DAISEP MabXpure is designed to trap various impurities
primarily targeting bioburden in the clarified cell harvest andbulk drug substance process intermediates. These impurities
and bioburden are considerably reduced or eliminated whilst
being retained by the resin and the mAbs are recovered in theflowthrough (Figure 1). The resin is packed in an integrated
format which has been designed to be a robust scalable plugand play device (Figure 2).
DAISEP MabXpure’s high retention capacity for impurities
and bioburden allows for high depletion rates of HCP andDNA >75% from the process feed/bulk drug substanceintermediates. MabXpure has a high selectivity for co-eluted
HCP traces, and simultaneously ensures high recovery ratesof the target molecules and mAbs (>95%), delivering a muchhigher quality drug substance or intermediate product in the
manufacturing process conventional and current purification
technologies.

Figure 2. MabXpure cartridge

WWW.DAICELBIOSEPARATIONS.COMDYNAMIC PERFORMANCE OF MABXPURE FTAPPLICATION NOTE #2

MATERIALS AND METHODSLOAD MATERIALBUFFER

CHO-K1 clarified feed,mAb 2g/L

DEVICETris HCl 50 mM pH 7.4

Depletion testing in the dynamic mode is performed using
the CHO-K1 process feed flow injection through MabXpure
cartridge. Collection of 5 column fractions (CVs) is performed
with different residence times. Following this flowthroughprocess, quantification of HCP is performed using the CHO-

MabXpure prepacked

specific ELISA kits from Cygnus and mAb titer is measured byHPLC-SEC. Additional pooled fraction analysis was performed
to compare the bioburden depletion with a 1:20 mix ratio (resinto product volumetric ratio) under static conditions.

RESULTS
In conventional AEX flowthrough applications, i.e., chromatography membranes and other AEX resins, the presence of divalent
phosphate ions in the feed limits the ion exchange mechanism, i.e., the adsorption capacity of the technology for acidic HCP andimpurities. However, there is a displacement effect observed after 20CVs where 75% HCPs are removed in the flowthrough andretained in the eluted fraction. Those performances of DAISEP MabXpure are consistent with the depletion levels observed understatic conditions (refer to Application Note #1).
In dynamic mode, the affinity for low molecular weight and negatively charged species increases at shorter residence times
where smaller impurities are forced to diffuse and penetrate into the porous structure of MabXpure and are trapped even more
effectively in comparison to static process conditions. MabXpure is ideally designed for significant depletion of HCP tracesfrom the process feed during the polishing stage. However, with DAISEP MabXpure, up to 75% of HCPs are removed under
flowthrough conditions (Figure 3, pool fraction – 20 CVs) and recovery yield is >95% (Figure 4).
HCP Depletion Performances vs residence timemAb Recovery Performances vs residence time

100100

98
90 89

96

100 100 98

100 100 99

10-15cvs

15-20cvs

94

95 96 93

76
75

5239

12 11 10Feed

1-5cvs2220 18 215-10cvs
1'

2210-15cvs
2'39

36

2515-20cvs

5'

Figure 3. Residual HCP with 3 different
residence times with DAISEP MabXpure.

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

25

Pool

Feed

1-5cvs5-10cvs
1'2'Pool

5'Figure 4. mAb recovery for 3 residence
times with DAISEP MabXpure.

WWW.DAICELBIOSEPARATIONS.COM

DYNAMIC PERFORMANCE OF MABXPURE FTAPPLICATION NOTE #2

DISCUSSION
In its pre-filled resin filter cartridge (RFC) design, the DAISEP
MabXpure fits very easily within a platform purification process
where flowthrough conditions need to be streamlined in theprocess following the clarification/harvest stage. Protecting theplatform whilst supplementing, complimenting or replacing theexisting chromatographic steps becomes easier like MabXpure
which is less intrusive to the product and the process eliminating
the need for a bind and elute process step operation.

DAISEP MabXpure can be easily applied to polish the process
feed without negatively impacting the process performance inthe presence of phosphate ions, and it does so ideally with ashort contact time. Its high throughput capacity is adequate
for sample preparation applications where a fast, simple androbust solution is desired for process developmental work orto find optimal conditions for in-process implementation ofMabXpure.

CONCLUSIONDAISEP MabXpure has a high potential of
depleting Host Cell Proteins and DNA withextremely high mAb recovery rates. Depending on
the process conditions and residence times, thecapacity of DAISEP MabXpure can eliminate up to
75% of HCPs with mAb recoveries >95%.

The simple design of DAISEP MabXpure(single-use pre-filled cartridge) makes it an ideal
flowthrough device to protect the downstream
platform as well as to polish the feed before andafter various chromatographic and TFF stages in
the purification process.

MabXpure can supplement, complement or replace
the pre-existing DSP unit operations

DAISEP MabXpure is easily scaleable and can beapplied at lab through to process GMP scale.

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

WWW.DAICELBIOSEPARATIONS.COM

STATICOFMABXPURE
DYNAMICPERFORMANCE OFFTSTATIC PERFORMANCEPERFORMANCEOFMABXPURE
MABXPURE#APPLICATIONAPPLICATION NOTE
NOTE #2# 11
APPLICATIONNOTEDAISEPDAISEP MABXPURE
MABXPURE RANGERANGEMABXPURE BULKMABXPURE BULK
MabXpure bulk 50 mLMabXpure bulk 50 mL
MabXpure bulk 500 mL
MabXpure bulk 500 mL
MabXpure bulk 1000 mL
MabXpure bulk 1000 mL
MabXpure bulk 10L
MabXpure bulk 10L

DMXBK0050
DMXBK0050
DMXBK0500
DMXBK0500
DMXBK1000DMXBK1000DMXBK10DMXBK10MABXPURE FTMABXPURE FT
MabXpure FT 1 mL (5 units/pack)
MabXpure FT 1 mL (5 units/pack)
MabXpure FT 5 mL (5 units/pack)
MabXpure FT 5 mL (5 units/pack)
MabXpure FT 50 mL (1 unit)
MabXpure FT 50 mL (1 unit)
MabXpure FT 500 mL (1 unit)
MabXpure FT 500 mL (1 unit)
MabXpure FT 1.5 L (1 unit)
MabXpure FT 1.5 L (1 unit)

DMXFT0001
DMXFT0001
DMXFT0005
DMXFT0005
DMXFT0050
DMXFT0050
DMXFT0500
DMXFT0500
DMXFT1500
DMXFT1500

MABXPURE KITMABXPURE KIT
MabXpure Kit
MabXpure Kit

DMXKT0001
DMXKT0001

50 mL
50 mL

500 mL
500 mL

1 mL
1 mL

5 mL
5 mL

MABXPURE KITMABXPURE KITDAISEP SPIN XS & XL
DAISEP SPIN XS & XL
DAISEP Spin XS (40 units/pack)
DAISEP Spin XS (40 units/pack)
DAISEP Spin XL (8 units/pack)
DAISEP Spin XL (8 units/pack)

DSPXS0040
DSPXS0040
DSPXL0008
DSPXL0008
XS

XL

XS

XL

DAICELDAICEL BIOSEPARATIONSBIOSEPARATIONS - LOCATIONSLOCATIONSNORTH/LATIN AMERICANORTH/LATIN
AMERICA
Chiral Technologies,
Inc.

CHINA
CHINA
Daicel Chiral Technologies (China) Co., Ltd

EUROPEEUROPE
Chiral Technologies Europe SAS

INDIA
INDIA
Daicel Chiral Technologies (India) Pvt. Ltd

ChiralTechnologies,
800 NorthFive PointsInc.
Road
800
North
FivePA
PointsRoadWest Chester,
19380
West
Chester,PA
19380
Tel: +1-610-594-2100
Tel: +1-610-594-2100
Fax:+1-610-594-2325
Fax:+1-610-594-2325
www.daicelbioseparations.com
www.daicelbioseparations.com
Email: bioseparations@cti.daicel.com
Email: bioseparations@cti.daicel.com

ChiralTechnologies
Europe SAS
Parc d’Innovation- 160,
Parc
d’Innovation
160,
Bd Gonthier d’Andernach - CS 80140
BdGonthier- CS 80140
67404
Illkirchd’Andernach
Cedex FRANCE
67404
Cedex
FRANCE
Tel:+33Illkirch(0) 3 88
79 5200
Tel:
Fax:+33+33(0)(0)33888879
665200
71 66
Fax:+33(0)
388
66 71 66
www.daicelbioseparations.com
www.daicelbioseparations.com
Email: bioseparations@cte.daicel.com
Email: bioseparations@cte.daicel.com

DaicelTechnologies(China) Co., Ltd
No. 32,Chiral
HeXiang
RoadNo.
32,HeXiang
Road
Waigaoqiao FreeTradeZone, Post code: 200131
Waigaoqiao
FreeTradeZone, Post code: 200131
Shainghai, China
Shainghai,
China60086
Tel:+86-21-504
Tel:Fax:+86-21-504+86-21-50460086
62321
Fax:+86-21-504 62321
www.daicelbioseparations.com
www.daicelbioseparations.com
Email: bioseparations@dctc.daicel.com
Email: bioseparations@dctc.daicel.com

DaicelTechnologies(India) Pvt. ParkLtdLab No:Chiral
A4, Phase
III, IKP Knowledge
Lab
No:A4,Phase
III,IKP
Knowledge
Park
Genome Valley, Turkapally, Shameerpet
GenomeValley,Turkapally, Shameerpet
Ranga Reddy
District
Ranga Reddy
District
Hyderabad- 500
078, Telangana INDIA
Hyderabad
500 3700
078, Telangana
INDIA
Tel: +91 40 -2338
or 91 40 2348
0103
Tel:+91402338
3700
or91402348
0103
Fax: +91 40 2348 0104
Fax:+914023480104
www.daicelbioseparations.com
www.daicelbioseparations.com
Email: bioseparations@dcti.daicel.com
Email: bioseparations@dcti.daicel.com

ASIAASIA
Daicel Corporation

DaicelCorporation
CPI Company
CPICompany
JR Shinagawa East Building
JRShinagawa
Building
2-18-1,Konan, EastMinato-ku
2-18-1,108-8230
Konan, Minato-ku
TokyoJAPANTokyo
108-8230
JAPAN
Tel: +81(0) 3 6711
8221
Tel:+81(0)36711
Fax: +81 (0) 3 67118221
8228
Fax:+81 (0) 3 6711 8228
www.daicelbioseparations.com
www.daicelbioseparations.com
Email: bioseparations@jp.daicel.com
Email: bioseparations@jp.daicel.com