Affinity Chromatography Resins - KANEKA KanCap™ · Affinity chromatography resins for full length...
Transcript of Affinity Chromatography Resins - KANEKA KanCap™ · Affinity chromatography resins for full length...
Affinity Chromatography Resins
CelluloseBase Matrix
KANEKA KanCap™ resins use a highly cross-
linked cellulose-based matrix with an average
particle of size 65 – 85 µm and reduced non-
specifi c binding properties when compared to
other base matrices such as polymers or glass.
The cellulose-based matrix is biocompatible and
has been successfully used for many years in
KANEKA’s plasmapheresis systems.
The combination of highly crosslinked cellulose-
based matrix with the
in-house engineered
ligands makes
KANEKA KanCap™
series an excellent
tool to effi ciently
capture and purify
high titer of antibodies
and derivatives. n
KANEKA offers two protein A affi nity
resins for industrial–scale purifi cation
of full length monoclonal antibodies:
KANEKA KanCapA™
& KANEKA KanCapA™ 3G.
Both resins are manufactured under
a validated GMP process to ensure
a stable and reliable supply chain.
The entire GMP production process is
free of animal derived components and
is BSE-TSE Free certifi ed. Regulatory
Support Files are available under a
Confi dential Disclosure Agreement (CDA).
Affi nity chromatography resins for full length antibody purifi cationKANEKA KanCap™ chromatography resins are effi cient cellulose-based
affi nity sorbents intended for purifi cation of all antibody formats. With
their innovative proprietary ligands, KANEKA KanCap™ series resins
demonstrate increased antibody affi nity and high binding capacity.
With these new resins Kaneka offers a comprehensive solution for
purifying all antibody formats from classical full-length antibodies
to Fc-fusion proteins, bispecifi c and fragmented formats:
Cellulose-based matrix microscope image (x20k).
series
1. KANEKA KanCapA™ 3G
is a Protein A resin with
enhanced performance.
2. KANEKA KanCapA™
is a Protein A resin
intended for industrial
scale purifi cation of mAb’s.
3. KANEKA KanCap™ L
is a Protein L resin
dedicated for purifi cation
of a wide range of antibody
fragments and derivatives
containing the κ VL
domain.
4. KANEKA KANCap™ G
is a Protein G resin
recommended for purifying
antibody fragments and
derivatives containing the
CH1 domain.
100µm
KANEKA KanCapA™ 3G’s ligand is a multimer of a modifi ed
C domain designed to improve binding capacity, elution profi le,
impurity removal ability and alkaline stability. n
KANEKA KanCapATM3G a resin with enhanced performance
1.
> Uniqueimpurity removal properties
> Excellent elution profi le
> Enhanced binding capacity
#2
Affinity chromatography resins for full length antibody purification
Performance
Enhanced binding capacity
Dynamic binding capacity (DBC)
is closely linked to productivity
and production costs. It is a
key parameter to consider
when selecting an efficient
chromatography resin.
KANEKA KanCapA™ 3G shows
enhanced binding capacity
which makes it suitable
for monoclonal antibody
purification from high titer
feedstocks. n DBC DEPENDENCY on the residence time (Polyclonal Human IgG).
ELUTION pH COMPARISON of mAbs and Fc fusion derivatives
3.4
3.6
A B C D E F G H
3.8
4.0
4.2
Elu
tio
n p
H
0
20
40
60
80
0 2 4 6 8 10
DB
C a
t 5
% b
reak
thro
ug
h (m
g lg
G /
mL
re
sin
)
Residence time (min.)
100
80
60
40
20
0
0 100 200 300
Cycle Number
Re
mai
nin
g B
ind
ing
Cap
acit
y [%
]
0.1 M NaOH
0.5 M NaOH
100
80
60
40
20
0
0 100 200 300
Cycle Number
Yie
ld [
%]
Yield
50
40
30
20
10
0
0 100 200 300
Cycle Number
Lig
and
Le
ach
ing
[p
pm
of
lgG
]
Ligand Leaching
800
700
600
500
400
300
200
100
0
0.00 0.05 0.10 0.15
Pressure [MPa]
Lin
ear
Flo
w R
ate
[cm
/h]
0.20 0.25 0.30
30 cm l.D. x 20 cm height
45 cm l.D. x 23 cm height
60 cm l.D. x 25 cm height
7 cm l.D. x 20 cm height
4.4 cm l.D. x 20 cm height
10 cm l.D. x 20 cm height
0
5
10
15
20
25
30
35
Bin
din
g C
ap
acit
y (m
g/m
L-g
el)
Fab(A) Fab(B) Fab(C)
Competitor A Competitor B
Fab(A) Fab(B) Fab(C)
KANEKA KanCapTM G
Fab(A) Fab(B) Fab(C)
3.4
3.6
A B C D E F G H
3.8
4.0
4.2
Elu
tio
n p
H
0
20
40
60
80
0 2 4 6 8 10
DB
C a
t 5
% b
reak
thro
ug
h (m
g lg
G /
mL
re
sin
)
Residence time (min.)
100
80
60
40
20
0
0 100 200 300
Cycle Number
Re
mai
nin
g B
ind
ing
Cap
acit
y [%
]
0.1 M NaOH
0.5 M NaOH
100
80
60
40
20
0
0 100 200 300
Cycle Number
Yie
ld [
%]
Yield
50
40
30
20
10
0
0 100 200 300
Cycle Number
Lig
and
Le
ach
ing
[p
pm
of
lgG
]
Ligand Leaching
800
700
600
500
400
300
200
100
0
0.00 0.05 0.10 0.15
Pressure [MPa]
Lin
ear
Flo
w R
ate
[cm
/h]
0.20 0.25 0.30
30 cm l.D. x 20 cm height
45 cm l.D. x 23 cm height
60 cm l.D. x 25 cm height
7 cm l.D. x 20 cm height
4.4 cm l.D. x 20 cm height
10 cm l.D. x 20 cm height
0
5
10
15
20
25
30
35
Bin
din
g C
ap
acit
y (m
g/m
L-g
el)
Fab(A) Fab(B) Fab(C)
Competitor A Competitor B
Fab(A) Fab(B) Fab(C)
KANEKA KanCapTM G
Fab(A) Fab(B) Fab(C)
DB
C a
t 5
% b
reak
thro
ug
h
Residence time (min.)
mA
b m
on
om
er
con
cen
tra
tio
n (
SE
C a
rea
)
mA
b m
on
om
er
con
cen
tra
tio
n (
SE
C a
rea
)
Re
lati
ve d
istr
ibu
tio
n o
f H
CP
(/%
)
Re
lati
ve d
istr
ibu
tio
n o
f a
gg
reg
ate
(%
)
Elu
tio
n p
H
80 4.2
4.0
3.8
3.6
3.4
60
40
20
2 4 6 8 10
0
16.000
KANEKA KanCapATM 3G KANEKA KanCapATM 3GCompetitor A Competitor A16.000
Fraction # Fraction #
12.000 12.000
8.000 8.000
4.000 4.000
0 0
0 A B C D E F G H
Excellent Elution pHKANEKA KanCapA™ 3G’s ligand
was designed to allow elution of
monoclonal antibodies and their
Fc fusion derivatives under very
mild acidic conditions, between
pH 3.7 and 4.2. n
Unique impurity removal properties
KANEKA KanCapA™ 3G is a new
generation of Protein A affinity
chromatography resins carrying a
recombinant protein A ligand with
enhanced properties. The resin
has a better potential for impurity
removal compared to current
commercially available resins. n
0
4,000
8,000
12,000
16,000
1 2 3 4 5 6 7 8 9 10 11
pH 3.8
0
20
40
60
80
1 2 3 4 5 6 7 8 9 10 11
pH 3.7
Aggregate removalKANEKA KanCapA™ 3G Competitor A
Fraction #
mA
b m
on
om
er
con
cen
trat
ion
(S
EC
are
a)
Re
lati
ve d
istr
ibu
tio
n o
f ag
gre
gat
e (
/%)
0
4,000
8,000
12,000
16,000
1 2 3 4 5 6 7 8 9 10 11
mA
b m
on
om
er
con
cen
trat
ion
(S
EC
are
a) pH 3.8
0
20
40
60
80
1 2 3 4 5 6 7 8 9 10 11
pH 3.7
Fraction #
Host cell protein removalKANEKA KanCapA™ 3G Competitor A
Re
lati
ve d
istr
ibu
tio
n o
f H
CP
(/%
)
HOST CELL PROTEIN (left)
AND AGGREGATES (right)
separation during mAb elution by linear
pH gradient.
Monomer Peak
Host cell protein (HCP)
Aggregate
Molecules: IgG1 (CCCF),
Load: 5 mg/mL resin.
Elution: 50 mM Citrate buffer,
(gradient pH from 4.5 to 3, 10 CV).
KANEKA KanCapATM 3G
Competitor A
KANEKA KanCapATM 3G Competitor A KANEKA KanCapATM
#3
Molecules: A,G, H = Humanized IgG1 B, F= Human IgG2 C = Fc-fusion D, E = Chimeric IgG
Elution: 50 mM citrate buffer gradient pH from 6 to 3.
1 2 3 4 5 6 7 8 9 10 11 1 2 3 4 5 6 7 8 9 10 111 2 3 4 5 6 7 8 9 10 11 1 2 3 4 5 6 7 8 9 10 11
0 0
20 20
40 40
60 60
80 80
Affinity Chromatography Resins
KANEKA KanCapA™’s ligand is a multimer of a modified C domain
containing amino acid mutations to improve the resins’ alkaline
stability and antibodies elution profile. n
KANEKA KanCapATM 2.
> Track record of successful mAb GMP manufacturing for clinical trial materials
> Alkaline stability and long lifetime
> High flow rate operation and easy to scale up from pilot to process
Performance
Alkaline stability and long lifetime
KANEKA KanCapA™ can be sanitized
with 0.1 or 0.5 M NaOH. It can be
safely used up to 300 cycles with a
limited loss of Binding Capacity after
alkaline CIP with 0.1 M NaOH for
15 minutes contact time.
Over the CIP cycles, the elution yields
and the levels of the leached Protein A
remain stable. n
3.4
3.6
A B C D E F G H
3.8
4.0
4.2
Elu
tio
n p
H
0
20
40
60
80
0 2 4 6 8 10
DB
C a
t 5
% b
reak
thro
ug
h (m
g lg
G /
mL
re
sin
)
Residence time (min.)
100
80
60
40
20
0
0 100 200 300
Cycle Number
Re
mai
nin
g B
ind
ing
Cap
acit
y [%
]
0.1 M NaOH
0.5 M NaOH
100
80
60
40
20
0
0 100 200 300
Cycle Number
Yie
ld [
%]
Yield
50
40
30
20
10
0
0 100 200 300
Cycle Number
Lig
and
Le
ach
ing
[p
pm
of
lgG
]
Ligand Leaching
800
700
600
500
400
300
200
100
0
0.00 0.05 0.10 0.15
Pressure [MPa]
Lin
ear
Flo
w R
ate
[cm
/h]
0.20 0.25 0.30
30 cm l.D. x 20 cm height
45 cm l.D. x 23 cm height
60 cm l.D. x 25 cm height
7 cm l.D. x 20 cm height
4.4 cm l.D. x 20 cm height
10 cm l.D. x 20 cm height
0
5
10
15
20
25
30
35
Bin
din
g C
ap
acit
y (m
g/m
L-g
el)
Fab(A) Fab(B) Fab(C)
Competitor A Competitor B
Fab(A) Fab(B) Fab(C)
KANEKA KanCapTM G
Fab(A) Fab(B) Fab(C)
3.4
3.6
A B C D E F G H
3.8
4.0
4.2
Elu
tio
n p
H
0
20
40
60
80
0 2 4 6 8 10
DB
C a
t 5
% b
reak
thro
ug
h (m
g lg
G /
mL
re
sin
)
Residence time (min.)
100
80
60
40
20
0
0 100 200 300
Cycle Number
Re
mai
nin
g B
ind
ing
Cap
acit
y [%
]
0.1 M NaOH
0.5 M NaOH
100
80
60
40
20
0
0 100 200 300
Cycle Number
Yie
ld [
%]
Yield
50
40
30
20
10
0
0 100 200 300
Cycle Number
Lig
and
Le
ach
ing
[p
pm
of
lgG
]
Ligand Leaching
800
700
600
500
400
300
200
100
0
0.00 0.05 0.10 0.15
Pressure [MPa]
Lin
ear
Flo
w R
ate
[cm
/h]
0.20 0.25 0.30
30 cm l.D. x 20 cm height
45 cm l.D. x 23 cm height
60 cm l.D. x 25 cm height
7 cm l.D. x 20 cm height
4.4 cm l.D. x 20 cm height
10 cm l.D. x 20 cm height
0
5
10
15
20
25
30
35
Bin
din
g C
ap
acit
y (m
g/m
L-g
el)
Fab(A) Fab(B) Fab(C)
Competitor A Competitor B
Fab(A) Fab(B) Fab(C)
KANEKA KanCapTM G
Fab(A) Fab(B) Fab(C)
3.4
3.6
A B C D E F G H
3.8
4.0
4.2
Elu
tio
n p
H
0
20
40
60
80
0 2 4 6 8 10
DB
C a
t 5
% b
reak
thro
ug
h (m
g lg
G /
mL
re
sin
)
Residence time (min.)
100
80
60
40
20
0
0 100 200 300
Cycle NumberR
em
ain
ing
Bin
din
g C
apac
ity
[%]
0.1 M NaOH
0.5 M NaOH
100
80
60
40
20
0
0 100 200 300
Cycle Number
Yie
ld [
%]
Yield
50
40
30
20
10
0
0 100 200 300
Cycle Number
Lig
and
Le
ach
ing
[p
pm
of
lgG
]
Ligand Leaching
800
700
600
500
400
300
200
100
0
0.00 0.05 0.10 0.15
Pressure [MPa]
Lin
ear
Flo
w R
ate
[cm
/h]
0.20 0.25 0.30
30 cm l.D. x 20 cm height
45 cm l.D. x 23 cm height
60 cm l.D. x 25 cm height
7 cm l.D. x 20 cm height
4.4 cm l.D. x 20 cm height
10 cm l.D. x 20 cm height
0
5
10
15
20
25
30
35
Bin
din
g C
ap
acit
y (m
g/m
L-g
el)
Fab(A) Fab(B) Fab(C)
Competitor A Competitor B
Fab(A) Fab(B) Fab(C)
KANEKA KanCapTM G
Fab(A) Fab(B) Fab(C)
RESIN LIFETIME EVALUATION after alkaline CIP with 0.1M NaOH for 15 min contact time. Remaining dynamic binding capacity at 5 % breakthrough (left); Yield (middle) and Protein A leaching (right). Ligand leaching: values less than the detection limit (1 ppm) are plotted as 1 ppm.
#4
100 100 100
80 80 80
60 60 60
40 40 40
20 20 20
0 0 0
Re
mai
nin
g B
ind
ing
Cap
acit
y (%
)
Cycle Number Cycle Number Cycle Number
Yie
ld (
%)
Lig
and
Le
ach
ing
(p
pm
of
IgG
)
0 100 200 300 0 100 200 300 0 100 200 300
0.1 M NaOH 0.5 M NaOH Yield Ligand Leaching
Mild acidic elution conditions for all mAbs
One of the most critical factors during monoclonal antibody
purifi cation is the pH used for their elution. In the case of
VH3-encoded mAbs, a low pH elution buffer is needed due
to their undesired binding to Protein A through the Fab region.
However it is well known that low pH elution leads to higher
aggregate formation.
To overcome this issue KANEKA KanCapA™ ligand has been
designed to remove the binding ability to Fabs. This allows milder
elution conditions. n
Operation at high fl ow rate and scalability from pilot to process
Thanks to its innovative highly crosslinked cellulose-based
matrix KANEKA KanCapA™ is perfectly suited for pilot and
large scale purifi cations. n
PRESSURE VS FLOW for packed columns of different diameters. The pressure generated by packed beds was calculated by substracting the pressure of the system from total pressure.
#5
ELUTION PROFILE of VH3-encoded mAb compared to other commercially available Protein A resins.Load: 5 g IgG /L resin; Strip solution: 1 M Acetic acid
pH 3.5pH 3.0
3.4
3.6
A B C D E F G H
3.8
4.0
4.2
Elu
tio
n p
H
0
20
40
60
80
0 2 4 6 8 10
DB
C a
t 5
% b
reak
thro
ug
h (m
g lg
G /
mL
re
sin
)
Residence time (min.)
100
80
60
40
20
0
0 100 200 300
Cycle Number
Re
mai
nin
g B
ind
ing
Cap
acit
y [%
]
0.1 M NaOH
0.5 M NaOH
100
80
60
40
20
0
0 100 200 300
Cycle Number
Yie
ld [
%]
Yield
50
40
30
20
10
0
0 100 200 300
Cycle Number
Lig
and
Le
ach
ing
[p
pm
of
lgG
]
Ligand Leaching
800
700
600
500
400
300
200
100
0
0.00 0.05 0.10 0.15
Pressure [MPa]
Lin
ear
Flo
w R
ate
[cm
/h]
0.20 0.25 0.30
30 cm l.D. x 20 cm height
45 cm l.D. x 23 cm height
60 cm l.D. x 25 cm height
7 cm l.D. x 20 cm height
4.4 cm l.D. x 20 cm height
10 cm l.D. x 20 cm height
0
5
10
15
20
25
30
35
Bin
din
g C
ap
acit
y (m
g/m
L-g
el)
Fab(A) Fab(B) Fab(C)
Competitor A Competitor B
Fab(A) Fab(B) Fab(C)
KANEKA KanCapTM G
Fab(A) Fab(B) Fab(C)
800
700
600
500
400
300
200
100
0
0.00 0.05 0.10 0.15 0.20 0.25 0.30
Pressure (MPa)
Lin
ear
Flo
w R
ate
(cm
/h)
KanCapAAgarosePolymerGlass
A280
VolumeELUTION STRIP
A280
ELUTION STRIP
A280
VolumeELUTION STRIP
A280
ELUTION STRIP
A280
Volume
Volume Volume Volume
ELUTION STRIP
A280
ELUTION STRIP
A2
80
KanCapAAgarosePolymerGlass
A280
VolumeELUTION STRIP
A280
ELUTION STRIP
A280
VolumeELUTION STRIP
A280
ELUTION STRIP
A280
Volume
Volume Volume Volume
ELUTION STRIP
A280
ELUTION STRIP
A2
80
KanCapAAgarosePolymerGlass
A280
VolumeELUTION STRIP
A280
ELUTION STRIP
A280
VolumeELUTION STRIP
A280
ELUTION STRIP
A280
Volume
Volume Volume Volume
ELUTION STRIP
A280
ELUTION STRIP
KanCapATM
AgarosePolymerGlass
3.4
3.6
A B C D E F G H
3.8
4.0
4.2
Elu
tio
n p
H
0
20
40
60
80
0 2 4 6 8 10
DB
C a
t 5
% b
reak
thro
ug
h (m
g lg
G /
mL
re
sin
)
Residence time (min.)
100
80
60
40
20
0
0 100 200 300
Cycle Number
Re
ma
inin
g B
ind
ing
Ca
pac
ity
[%]
0.1 M NaOH
0.5 M NaOH
100
80
60
40
20
0
0 100 200 300
Cycle Number
Yie
ld [
%]
Yield
50
40
30
20
10
0
0 100 200 300
Cycle Number
Lig
an
d L
ea
chin
g [
pp
m o
f lg
G]
Ligand Leaching
800
700
600
500
400
300
200
100
0
0.00 0.05 0.10 0.15
Pressure [MPa]
Lin
ear
Flo
w R
ate
[cm
/h]
0.20 0.25 0.30
30 cm l.D. x 20 cm height
45 cm l.D. x 23 cm height
60 cm l.D. x 25 cm height
7 cm l.D. x 20 cm height
4.4 cm l.D. x 20 cm height
10 cm l.D. x 20 cm height
0
5
10
15
20
25
30
35
Bin
din
g C
ap
acit
y (m
g/m
L-g
el)
Fab(A) Fab(B) Fab(C)
Competitor A Competitor B
Fab(A) Fab(B) Fab(C)
KANEKA KanCapTM G
Fab(A) Fab(B) Fab(C)
3.4
3.6
A B C D E F G H
3.8
4.0
4.2
Elu
tio
n p
H
0
20
40
60
80
0 2 4 6 8 10
DB
C a
t 5
% b
reak
thro
ug
h (m
g lg
G /
mL
re
sin
)
Residence time (min.)
100
80
60
40
20
0
0 100 200 300
Cycle Number
Re
ma
inin
g B
ind
ing
Cap
acit
y [%
]
0.1 M NaOH
0.5 M NaOH
100
80
60
40
20
0
0 100 200 300
Cycle Number
Yie
ld [
%]
Yield
50
40
30
20
10
0
0 100 200 300
Cycle Number
Lig
an
d L
ea
chin
g [
pp
m o
f lg
G]
Ligand Leaching
800
700
600
500
400
300
200
100
0
0.00 0.05 0.10 0.15
Pressure [MPa]
Lin
ear
Flo
w R
ate
[cm
/h]
0.20 0.25 0.30
30 cm l.D. x 20 cm height
45 cm l.D. x 23 cm height
60 cm l.D. x 25 cm height
7 cm l.D. x 20 cm height
4.4 cm l.D. x 20 cm height
10 cm l.D. x 20 cm height
0
5
10
15
20
25
30
35
Bin
din
g C
ap
acit
y (m
g/m
L-g
el)
Fab(A) Fab(B) Fab(C)
Competitor A Competitor B
Fab(A) Fab(B) Fab(C)
KANEKA KanCapTM G
Fab(A) Fab(B) Fab(C)
4.4 cm l.D. x 20cm height
7 cm l.D. x 20cm height
10 cm l.D. x 20cm height
30 cm l.D. x 20cm height
45 cm l.D. x 20cm height
60 cm l.D. x 20cm height
SCHEMATIC REPRESENTATION of KANEKA KanCapTM L’s ligand affi nity to different kappa light chain of different antibody fragments in comparison to wild type protein L.
Affi nity
Fab Variants
Affi nity
Fab Variants
Wild type Protein L KANEKA KanCapTM L Protein L
KANEKA offers also TWO solutions
for purifying antibody fragments
and derivatives that lack the Fc region:
KANEKA KanCap™ L and
KANEKA KanCap™ G.
Affi nity chromatography resins for antibody fragments and derivatives purifi cation
KANEKA KanCap™ L is a newly developed cellulose-based Protein L
resin designed for capture and purifi cation of a wide range of antibody
fragment formats such as scFv, Fab and F(ab’)2. KANEKA KanCapTM L
ligand was designed to bind a wide spectra of molecules and to exibit
high affi nity to the VL domain of the antibody kappa light chain. It
is also an alternative tool for purifying full length antibody formats
containing a kappa light chain which poorly bind to Protein A resins. n
KANEKA KanCapTM L 3.
> High binding capacity for antibodies containing a κ light chain
> Wide binding range for antibody molecules containing a κ light chain FULL LENGTH ANTIBODY
Fc
Fab
Heavy Chain
Constant Region
Variable Region
FULL LENGTH ANTIBODY
Heavy Chain
#6
Affi nity Chromatography Resins
DBCSBC
DBCSBC
Fab (A) from humanized IgG1 kappa, Fab (B) from chimeric IgG1 kappa
Bin
ding
Cap
acit
y (m
g/m
L)
0
10
20
30
40
50
60
70
CompetitorKANEKA KanCapTM L
DBCSBC
DBCSBC
Fab (A) from humanized IgG1 kappa, Fab (B) from chimeric IgG1 kappa
Bin
ding
Cap
acit
y (m
g/m
L)
0
10
20
30
40
50
60
70
CompetitorKANEKA KanCapTM L
1 min. 4 min.
Fab A Fab B Fab A Fab B Fab A Fab B Fab A Fab B
4 min. 4 min. 4 min.1 min. 1 min. 1 min. 1 min. 4 min. 4 min. 4 min. 4 min.1 min. 1 min. 1 min.
Fab (A) from humanized IgG1 kappa, Fab (B) from chimeric IgG1 kappa
KANEKA KanCapTM L Competitor
DYNAMIC AND STATIC binding capacity evaluation.Fab (A) from humanized IgG1 kappa; Fab (B) from chimeric IgG1 kappa
70 70
60 60
50 50
40 40
30 30
RT 1 min. RT 4 min. SBC RT 1 min. RT 4 min. SBC
20 20
10 10
0 0
Bin
din
g C
ap
aci
ty (
mg
/mL
)
Fab A Fab B
Fab A Fab B
VH
CH1
CL
KANEKAKanCapTM G KANEKA
KanCapTM L
VL
KANEKA KanCap
VL
G
Light Chain
n KANEKA and the KANEKA logo are trademarks of
KANEKA.n KANEKA KanCapA™ is a
trademark of KANEKA.n KanCap™ is a trademark
of KANEKA.n RoboColumn® is a
registered trademark of Repligen GmbH.
n KANEKA products may not be resold or
transferred, modifi ed for resale or transfer, or used
to manufacture commercial products for these purposes
without written approval from KANEKA.
n The information in this brochure were obtained by
KANEKA CORPORATION.n Disclaimers: All
experimental data are provided “as is”, without
any warranty of accuracy or completeness.
©2018 KANEKA CORPORATION, JAPAN
All rights reserved.
Affi nity chromatography resins for antibody fragments and derivatives purifi cation
KANEKA KanCapTM G is an innovative cellulose-based affi nity
resin designed for effi cient capture and purifi cation of antibody
fragment formats containing the human CH1 domain such as Fab
and F(ab’)2. KANEKA KanCapTM G ligand exhibits an increased
binding affi nity for the CH1 domain. It can also be used to purify full
length antibody formats. n
KANEKA KanCapTM G 4.
> High binding capacity for antibody molecules containing CH1 or Fc region
series
#7
CH1
VHVL
CL
CH1
VHVL
CL
Low affi nity
High affi nity
KANEKA KanCapTM G Protein G
Wild type Protein G
SCHEMATIC REPRESENTATION of KANEKA KanCapTM G’s ligand affi nity to CH1 domain
SDS-PAGE analysis of non-reduced Fab samples purifi ed from yeast supernatant M. Marker; 1.Supernatant; 2.Flow-through; 3.Wash; 4.Eluate
DYNAMIC BINDING capacity evaluation at 4 min. residence time.Fab (A) from monoclonal chimeric IgG1
Fab (B) from polyclonal human IgG (containing κ and λ types)
Fab (C) from monoclonal humanized IgG1
3.4
3.6
A B C D E F G H
3.8
4.0
4.2
Elu
tio
n p
H
0
20
40
60
80
0 2 4 6 8 10
DB
C a
t 5
% b
rea
kth
rou
gh
(mg
lgG
/ m
L r
esi
n)
Residence time (min.)
100
80
60
40
20
0
0 100 200 300
Cycle Number
Re
mai
nin
g B
ind
ing
Cap
acit
y [%
]
0.1 M NaOH
0.5 M NaOH
100
80
60
40
20
0
0 100 200 300
Cycle Number
Yie
ld [
%]
Yield
50
40
30
20
10
0
0 100 200 300
Cycle Number
Lig
and
Le
ach
ing
[p
pm
of
lgG
]
Ligand Leaching
800
700
600
500
400
300
200
100
0
0.00 0.05 0.10 0.15
Pressure [MPa]
Lin
ear
Flo
w R
ate
[cm
/h]
0.20 0.25 0.30
30 cm l.D. x 20 cm height
45 cm l.D. x 23 cm height
60 cm l.D. x 25 cm height
7 cm l.D. x 20 cm height
4.4 cm l.D. x 20 cm height
10 cm l.D. x 20 cm height
0
5
10
15
20
25
30
35
Bin
din
g C
ap
acit
y (m
g/m
L-g
el)
Fab(A) Fab(B) Fab(C)
Competitor A Competitor B
Fab(A) Fab(B) Fab(C)
KANEKA KanCapTM G
Fab(A) Fab(B) Fab(C)
KANEKA KanCapTM G Competitor A Competitor B
0
5
10
15
20
25
30
35
Fab(A) Fab(A) Fab(A)Fab(B) Fab(B) Fab(B)Fab(C) Fab(C) Fab(C)
Bin
din
g C
apac
ity
(mg
/mL
-ge
l)
JAPAN Kaneka Corporation1-12-32, Akasaka, Minato-ku,Tokyo 107-6028, [email protected]
USAKaneka US Innovation Center7979 Gateway Blvd., Suite 220Newark, CA [email protected]
EUROPEKaneka Eurogentec S.A.Rue du bois Saint-Jean, 144102 Seraing, [email protected]
www.bioseparation.kaneka.com
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