KANEKA KanCapATM Prepacked Column

i90~210¹ Sizej
Code No. 11101081 (1 mL)
11701083 (5 mL)
11501084 (5 mL~5)
For Genetic Research
KANEKA KanCapATM Prepacked Column
KANEKA KanCapATM Prepacked Column is a prepacked
column with KANEKA KanCapATM. KANEKA KanCapATM is
composed of highly crosslinked cellulose beads and Alkaline
Resistant Protein A ligand, which was designed to purify
immunoglobulins and Fcchimeric proteins from biological sam
ples and cell culture media. KANEKA KanCapATM Prepacked
Column can be used for screening work, initial stage process
development, and small scale purification.
Features
1. Alkaline Resistance and Long Life Cycle
2. Optimized Elution pH
3. High Binding Capacity
4. High Flow Rate Operation and Easy Scale Up from Pilot to
largescale Process
5. Prepacked Column Type and Ready to Use
kAbout Columnl
E Components of Prepacked Column are made from biocom
patible polymers, mainly highdensity polyethylene (column
tube, Lock nut, filter adapter). Filter plates are made from
porous polypropylene/polyethylene material.
E KANEKA KanCapATM Prepacked Column can be reused and
are not designed to be repacked.
E KANEKA KanCapATM Prepacked Column can be connected
directly to most liquid chromatography systems or worksta
tions. The connection to earlier FPLCR system is simple with
standard 1032 male/M6 female adapters.
E KANEKA KanCapATM Prepacked Column can also be oper
ated with syringes or peristaltic pumps.
kSafety Precautionsl
E Keep away from fire. This product is shipped as prepacked in
20% ethanol solution.
E Wear eye protection, protective gloves and lab coat.
E Make sure to tighten the connector to prevent leakage.
E During operation, make sure not to exceed pressure limit of
the column and pump system.
E Do not disassemble the product.
kProduct Descriptionl
Table 1 : Characteristics of KANEKA KanCapATM Prepacked
Column
Column Type
Inner Diameter~
Length (mm~mm)
Column Cross Section
(cm2)
Column Volume (mL)
Recommended Flow
rate1) (mL/min/cm/h)
Max. Flow rate
(mL/min/cm/h)
Outer Dimensions2)
(mm~mm)
Connectors
Maximum Operating
Pressure
Chemical Stability3)
Storage Condition4)
1)
2)
3)
4)
Lock nut
Column
Information
Column Tube
Lock nut
Outlet stopper
Fig. 1 Parts of the KANEKA KanCapATM Prepacked Column
(Code No. 11101081, 1mL shown)
| 1/8 |
5 mL (Code No.
11701083, 11501084)
8.0~100
0.5
0.5
1.0
0.170.25/2030
5.0
0.841.26/100150
0.84/100
4.19/500
10~84
10~164
Standard HPLC (1032 for 1/16 inch capillaries)
30 bar
30 bar
Column are tolerant to aqueous buffers and salt
solutions, 1M alkaline solutions, 1M nonoxidiz
ing mineral acids, 8M urea, 6M guanidine
hydrochloride, nonhalogenated organic solvents
and detergents. They are not compatible with
strong oxidants and halogenated solvents.
20% ethanol at 2`10ú
Recommended residence time is 4`6 min.
Including stoppers.
The chemical stability refers to the column hardware parts only. For
the chemical stability of KANEKA KanCapATM, please see ``Table
2 : Physical and chemical properties of KANEKA KanCapATM''.
Prevent from freezing.
Table 2 : Physical and chemical properties of KANEKA KanCapATM
Base Matrix
Inlet stopper
1 mL (Code No.
11101081)
8.0~20
Highly crosslinked cellulose
Average Particle Size1') 6585Êm
Ligand
Recombinant Protein A (Alkaline Resistant)
Coupling Chemistry
Reductive amination
Dynamic Binding
35 mg human polyclonal IgG/mL packed
Capacity2')
Chemical Stability3')
Working pH Range3')
resin
Stable in commonly used solvents in affinity
chromatography
pH 2pH13
CIP (Cleaning In Place) 0.10.5 mol/L sodium hydroxide, 68 mol/L
condition3')
urea, 6 mol/L guanidine hydrochloride
Operational Flow Rate
Up to 500 cm/h (bed height : 20`25 cm)
Residence Time
Z3 min (4`6 min. is recommended)
Storage Condition4')
Slurry in 20% ethanol at 2`10ú
| 2/8 |
1') Average particle size is medium particle size of the cumulative
volume distribution.
2') 5% dynamic binding capacity is determined by frontal analysis at 3
minutes of residence time.
3') Alkaline conditions can be used as CIP but are unsuitable for long
term exposure.
4') Prevent from freezing.
kOperationl
1. Connecting Column
1) Loosen the Outlet stopper (Do not over loosen).
2) Remove the Inlet stopper and connect the column to the
inlet line by droptodrop¦1 to avoid introducing air into
the column.
¦ 1 c Droptodrop means that the column is connected
to the inlet line as liquid and liquid are touching
with the liquid accumulated in connection.
3) Immediately remove the Outlet stopper and connect the
column to the outlet line.
2. Chromatographic Conditions
An example of chromatographic condition is described
below. The optimal condition depends on the characteristics
of feed material.
Flow rateÛ) :
Equilibration :
Load :
WashÜ) :
ElutionÝ) :
Strip :
NeutralizationÞ) :
CIPß) :
Reequilibration :
Storageà) :
2030 cm/h (1 mL type), 100150 cm/h (5 mL
type)
PBS, pH 7.4 (5 column volumes (CV))
IgG solution pH neutralized
PBS, pH 7.4 (5 CV)
Acetate buffer or Citrate buffer, pH3.5
1M acetic acid (3 CV)
PBS, pH 7.4 (3 CV)
0.1 M sodium hydroxide (contact time 15 min)
PBS, pH 7.4 (5 CV)
20% ethanol (5 CV)
Û) Recommended residence time is 4`6 min. The flow rate
corresponding with 4`6 min. of residence time depends on
column length. For 5 mL type, 100150 cm/h gives 4`6 minutes
of residence time.
Ü) Intermediate wash is conducted after washing step, if necessary.
Ý) Low pH elution may leads to higher aggregate formation of sam
ple. KANEKA KanCapATM can elute IgGs at mild acidic pH.
The eluted fraction is neutralized by adding a small amount of
neutralizing buffer (e.g. Tris buffer).
Þ) Direct contact of acid solution and alkaline solution on column
should be avoided.
ß) 0.5 M sodium hydroxide can also be used (see, ``CleaningIn
Place (CIP)''). Column should be neutralized after CIP.
à) See ``Storage''.
kCleaningInPlace (CIP)l
During the purification process, some contaminants, which
negatively affect the overall performance of the affinity chro
matography, might be accumulated in the column. It may
increase back pressure or reduce binding capacity. CIP can be
| 3/8 |
performed periodically to remove such contaminants and wash
out the column. Recommended CIP solution is 0.1 to 0.5 M sodi
um hydroxide. However, longer exposure time and higher con
centration of sodium hydroxide should be avoided to minimize
the loss of binding capacity. 68 M urea and 6 M guanidine
hydrochloride can also be applied. 70% ethanol can be used to
remove lipids. Different CIP protocols may have to be combined
depending on the contaminants to be removed.
kSanitizationl
KANEKA KanCapATM Prepacked Column can be sanitized by
washing with 0.1 to 0.5 M sodium hydroxide. However, longer
exposure time and higher concentration of sodium hydroxide
should be avoided to minimize the loss of binding capacity. 70%
ethanol also can be used.
kStoragel
To maintain a good column performance and prevent microbial
growth, unused KANEKA KanCapATM Prepacked Column
should be stored at 2 ` 10 ú in 20% ethanol. DO NOT FREEZE.
2% benzyl alcohol could also be used as storage buffer at 4 ú.
Remove KANEKA KanCapATM Prepacked Column from
system and seal the column by tightening both Inlet stopper and
Outlet stopper. Avoid introducing air into the column.
Wako Pure Chemical Industries, Ltd.
12, Doshomachi 3Chome, ChuoKu, Osaka 5408605, Japan
Telephone : {81662033741
Facsimile : {81662015964
http://www.wakochem.co.jp
Wako Chemicals USA, Inc.
Wako Chemicals GmbH
1600 Bellwood Road
Richmond, VA 23237
U.S.A.
Telephone : {18042717677
Facsimile : {18042717791
http://www.wakousa.com
Fuggerstrasse 12
D41468 Neuss
Germany
Telephone : {4921313110
Facsimile : {492131311100
http://www.wakochemicals.de
| 4/8 |
R[h No. 11101081i1 mLj
11701083i5 mLj
11501084i5 mL~5j
â`q¤p
KANEKA KanCapATM vpbNJ
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ters ð²gpº³¢B
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k»iTvl
Table 1 : KANEKA KanCapATM vpbNJÌÁ¥
1 mL (Code No.
5 mL (Code No.
11101081)
11701083, 11501084)
8.0~100
àa~·³(mm~mm) 8.0~20
0.5
0.5
JfÊ (cm2)
1.0
5.0
JeÊ (mL)
0.170.25/2030
0.841.26/100150
§¬¬1)
(mL/min/cm/h)
0.84/100
4.19/500
Åå¬¬
(mL/min/cm/h)
10~164
O¡@2) (mm~mm) 10~84
Standard HPLC (1032 for 1/16 inch capillaries)
RlN^[
30 bar
30 bar
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J^Cv
1) Ø¯ÔÍA4 ` 6 ªÅ·B
2) stoppersðÜÝÜ·B
3) JiÌ»wÀè«îñÅ·BKANEKA KanCapATM Ì»w
Àè«ÉÂ¢ÄÍATable 2 : KANEKA KanCapATM Ì¨IyÑ
»wI«¿ð²QÆº³¢B
4) µÈ¢Å¾³¢B
Table 2 : KANEKA KanCapATM Ì¨IyÑ»wI«¿
}gbNX
½Ï±qa
Lock nut
Column
Information
Kh
veCACg·¦ÌiAJÏ«j

»wiÒ³IA~m»j

®IeÊ
»wÀè«
Column Tube
Lock nut
Outlet stopper
} 1 KANEKA KanCapATM vpbNJÌp[c
i8mm I.D.~20mm heightj
| 5/8 |
Ë´Z[X
6585Êm
1')
pHÍÍ
ÅÀè
pH 213
3')
3')
0.10.5 mol/L_»igE
68 mol/LAfA6 mol/L_OAjW
Åå500 cm/hiwF2025 cmj
¬
Ø¯Ô
Û¶ð
35 mg qg|N[iIgG/mLSÌ
AtBjeB[N}gOtB[Äpn}
3')
èuôòð
¬
2')
Z3 ªi§F46ªÔj
4')
20%G^m[÷tC2`10ú
| 6/8 |
1') ½Ï±qaÍAÝÏÌÏªzÌÔ±aÅ·B
2') 5 ÷®IeÊÍO[ªÍiØ¯Ô 3 ªjÅªèµÄ¢Ü·B
3') AJ«ðÅôòÅ«Ü·ªA·úÔÌôòìÍð¯Ä¾
³¢B
4') µÈ¢Å¾³¢B
kìTvl
1. JÌÚ±
1) Outlet stopper ðÉßéBiÉß·¬È¢Å¾³¢Bj
2) Inlet stopper ðOµÄAJÉóCªüéÌðð¯é
½ßÉA Drop to Drop¦1 ÅüCÉJðÚ±
·éB
¦ 1 cÚ±Étð½ßÄAtÆtðGêí¹éæ
¤ÉÚ±·éB
3) Ú±ã¼®É Outlet stopper ðOµÄoCÉÚ±
·éB
2. N}gOtð
N}gOtðÌQláðºLÉ¦µÜ·BÅKÈð
ÍÞ¿Ì«¿ÉË¶µÜ·ÌÅ²¢º³¢B
¬¬Û)
½t»
Tv
ôòÜ)
noÝ)
Xgbv
aÞ)
CIPß)
Ä½t»
ÛÇà)
2030 cm/h(1 mL^Cv)A100150 cm/h(5 mL
^Cv)
PBS, pH 7.4(5 column volumes(CV))
pHðaµ½IgGnt
PBS, pH 7.4(5 CV)
|_obt@[Ü½ÍNG_obt@[,
pH 3.5
1 M|_(3 CV)
PBS, pH 7.4(3 CV)
0.1 M_»igE(ÚGÔ15ª)
PBS, pH 7.4(5 CV)
20÷G^m[(5 CV)
§·é CIP ntÍ 0.1 ©ç 0.5 mol/L _»igEÅ
·ªA·ÔÌ\IâZxÅÌ²gpÍAeÊðáº³
¹é°êª é½ßð¯Ä¾³¢BÜ½A 68 mol/L Afâ
6 mol/L _OAjWà CIP ntÆµÄgpÅ«A 70 ÷G^
m[Í¿ÉgpÅ«Ü·B±Ìæ¤ÈntðgÝí
¹é±ÆÉæÁÄlXÈ¬ü¨¿ÉÎÅ«Ü·B
kEÛ@l
KANEKA KanCapATM vpbNJÍA 0.1 ` 0.5
mol/L _»igEÅôò·é±ÆÅEÛÅ«Ü·Bµ©
µÈªçA·ÔÌ\IâZxÅÌ²gpÍAeÊðá
º³¹é°êª é½ßð¯Ä¾³¢B 70 ÷G^m[àg
pÅ«Ü·B
kÛ ¶l
J«\ðÛA÷¶¨Bðh®½ßA¢gpÌ
KANEKA KanCapATM vpbNJÍA 20 ÷G^m[
A 2 ` 10 úÅÛ¶µÄº³¢B{iÍAsÂÅ·B 2 ÷
xWAR[à 4 úÉ¨¯éÛÇpÉÕtÆµÄgpÅ
«Ü·B
VXe©çæèOµ½ KANEKA KanCapATM vpbN
JÌ¼[ð Inlet stopper Æ Outlet stopper Å÷ßÄÛÇ
µÄº³¢B»ÌÛAóCªüçÈ¢æ¤ÉCðÂ¯Ä¾³
¢B
Û) §Ø¯ÔÍA4 ` 6 ªÅ·B4 ` 6 ªÌØ¯ÔÉêv·
é¬¬ÍJÌ·³ÉË¶ÉµÜ·B5 mLÌJÌêA
100150 cm/h Ì¬¬Å 4 ` 6 ªÌØ¯ÔÅ·B
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¢B
Ý) ápHÅÌnoÍAÃWÌ`¬ðø«N±·©àµêÜ¹ñB
KANEKA KanCapATM Í}ChÈpHðÅIgGsðnoÅ
«Ü·BnoãÍTrisobt@[ðnotNVÉYÁ
µÄaìÆðsÁÄ¾³¢B
Þ) _ntâAJntÌJãÅÌ¼ÚIÚGÍð¯Ä¾
³¢B
ß) 0.5 M _»igEàgpÅ«Ü·B CleaninhInPlace
(CIP)ð²QÆº³¢BCIPãÍJðaµÄ¾³¢B
à) Û¶ð²QÆº³¢B
kCleaningInPlace (CIP)l
¸»HöÉ¨¢ÄAAtBjeB[N}gOtB[«\
SÌÉ«e¿ðyÚ·æ¤È¬ü¨¿ªJÉ~Ï·é°ê
ª èÜ·B»Ìæ¤È¨¿ÍAw³ðã¸³¹A\ðá
º³¹éÂ\«ª èÜ·B CIP Í»Ìæ¤È¬ü¨¿ðèú
IÉæè«AJðôò·é½ßÉÀ{³êÜ·B
STUVW XYZ[\
| 7/8 |
| 8/8 |
1612K01

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