Asahi Kasei - Qyu speed d aex Bioprocess Innovation

1. QyuSpeed D
AEX chromatography membrane,
a highly cost effective technology
5th ANNUAL BIO INNOVATION
15-16th February 2012
London UK
Bixente MARTIRENE – MSc Chem. Eng.
Product Manager
Asahi Kasei Bioprocess Europe

2. Content
PART 1 – Asahi Kasei Bioprocess
PART 2 – QyuSpeed D
PART 3 – Membrane vs. Resin
PART 4 – Performances
PART 5 – Cost simulation: Q resin,
Single Use Q
membrane,
QyuSpeed
D

3. PART 1 – Asahi Kasei Bioprocess

4. Asahi Kasei Group
HQ: Tokyo; Foundation: 1931; Employees: 25 000; Turnover: 16 B$
Asahi Kasei Medical
BIOPROCESS
Leukocyte Reduction (SepacellTM)
4

5. Asahi Kasei Bioprocess
 For 20 years, leader in virus removal
filtration with Planova filters range.
From 4.0 m2 down to 0.001 m2 surface area
5

6. Asahi Kasei Bioprocess
 For 20 years, leader in virus removal
filtration with Planova filters range.
 From 2010, new separation and purification
solutions, for biopharmaceuticals production.
 New products based on Asahi’s core
technology (hollow fibers) and high quality
standards.
6

7. Membranes for USP and DSP
7

8. PART 2 – QyuSpeed D

9. Specifications
Filter Format Hollow Fiber
Ligand type Anion exchange
Ligand DEA : -N+H-(CH2CH3)2
Pore Size 0.2 - 0.3 µm
Membrane Material Polyethylene
Fiber ID / OD 2.2 mm / 3.6 mm
Operating Pressure < 2 bar
Reusable after regeneration Yes
Outlet
Inlet
Drain
Hollow fiber 9

10. Pore structure and ligand configuration
Poly (glycidyl methacrylate)
Chain (backbone of the grafted chain)
micropore
)
H3C- C-C-O-CH2CHCH2
=
H 2C O HO N(CH2CH3)2
) (
H3C- C-C-O-CH2CHCH2
=
O HO N(CH2CH3)2
H 2C
(
Micropore
Nominal pore size: Grafted chain Anion-exchange group (DEA)
0.2-0.3 µm
Flow
direction Ligand density: 0.55-0.6 mmol/mL-ads
1200um
SEM x25 10

11. Separation mechanism
 Selectivity by ligand type.
 Grafted chains Multipoint adsorption
High Dynamic Binding Capacity (DBC).
Protein solution
Proteins - Proteins +
Convective flow
Grafted chain
with Ligands
Hollow Fiber
Pore of the membrane
11

12. Application
Removal of impurities from protein/MAb solutions:
DNA, Viruses, HCP, Prion, Endotoxin.
Host cell protein
Endotoxin
Isoelectric
DNA Viruses
Point
2 7 MAbs 12
Binding of Flow through of
impurities - pH protein/MAb +
buffer
12

13. Product line
Lab-filter
Membrane Volume 0.6 mL
Max Flow rate 8 mL/min
Availability From 2010
2-inch-filter
Membrane Volume 150 mL
Max Flow rate 2 L/min
Availability Mid 2011
4-inch-filter
Membrane Volume 550 mL
Max Flow rate 8 L/min
Availability End 2011
QyuSpeedD-5L end 2012!
315 mm
13

14. Where implementing QyuSpeed D ?
 Before or after Protein A.
 High salt tolerance = possible after CEX
without any prior dilution.
Centri- AIEX
Depth
fugation Filtration
or
Protein A
0.2 µm Planova UF
filtration
CIEX
Tangential MF
14

15. Where implementing QyuSpeed D ?
After cell culture, 3 actions in 1 single step:
 Cell culture clarification,
 AEX chromatography,
 “Protection” of Protein A.
Centri- AIEX
Depth
fugation Filtration
or
Protein A
0.2 µm Planova UF
filtration
CIEX
Tangential MF
15

16. PART 3 – Membrane vs. Resin

17. Membrane vs. Resin
 Convective vs. diffusive mass transfer.
 DBC independent of the flow rate.
 5-20 MV*/min vs. 0.5-2 CV**/min.
 Very high flow rates w/o high pressure drop.
*MV: Membrane Volume
**CV: Column Volume
Electron micrograph Electron micrograph
of QyuSpeed D of resin beads
17

18. Membrane vs. Resin
Resin columns can be oversized because :
slow binding mechanism by diffusion,
bed height limitation (~ 20-30 cm) to avoid too
high pressure drop,
poor salt tolerance,
lower impurities DBC than claimed BSA DBC.
18

19. Membrane vs. Resin
 To purify the same protein solution:
Membrane Volume (MV) << Column Volume (CV)
 MAb loading capacity: often > 2 000 g/L-QSD
19

20. PART 4 – Performances

21. Main AEX membranes - Performances
Q resin Membrane 1 Membrane 2 Membrane 3 QyuSpeed D
Configuration Beads Rolled Sheet Open Pleated Stacked Flat Sheet Hollow fibers
Medium Material Agarose Cellulose PES PE PE
-N+-(CH3)3 -N+-(CH3)3 Quaternary -N+H-
Ligand Primary amine
amine (CH2CH3)2
Max Flow rate 1 CV/min 30 MV/min 10 MV/min 12.5 MV/min 13.5 MV/min
Pore Size - > 3 µm 0.8 µm 0.65 µm 0.2-0.3 µm
10% BSA DBC
> 100 > 29 > 60 > 50 > 40
(g/L-medium)
10% DNA DBC
<5 < 10 < 35 < 35 < 35
(g/L-medium)
at ~ 0 mS/cm
10% DNA DBC < 30
Not applicable.
<2 < 30 < 30
(g/L-medium) (if another Max 5 mS/cm
type)
at 15 mS/cm
Virus LRV > 3-4 > 3-4 > 3-4 >4 >4
Usually much lower reduction than for DNA and Viruses. Very much depending on the operating
HCP reduction
conditions and HCP characteristics (large pI range).
Number of
Up to 100 Single use Not precised Single use > 10
regenerations
21

22. Main AEX membranes - Performances
Q resin Membrane 1 Membrane 2 Membrane 3 QyuSpeed D
Configuration Beads Rolled Sheet Open Pleated Stacked Flat Sheet Hollow fibers
Medium Material Agarose Cellulose PES PE PE
-N+-(CH3)3 -N+-(CH3)3 Quaternary -N+H-
Ligand Primary amine
amine (CH2CH3)2
Max Flow rate 1 CV/min 30 MV/min 10 MV/min 12.5 MV/min 13.5 MV/min
Pore Size - > 3 µm 0.8 µm 0.65 µm 0.2-0.3 µm
10% BSA DBC
> 100 > 29 > 60 > 50 > 40
(g/L-medium)
10% DNA DBC
<5 < 10 < 35 < 35 < 35
(g/L-medium)
at ~ 0 mS/cm
10% DNA DBC < 30
Not applicable.
<2 < 30 < 30
(g/L-medium) (if another Max 5 mS/cm
at 15 mS/cm type)
Virus LRV > 3-4 > 3-4 > 3-4 >4 >4
Usually much lower reduction than for DNA and Viruses. Very much depending on the operating
HCP reduction
conditions and HCP characteristics (large pI range).
Number of
Up to 100 Single use Not precised Single use > 10
regenerations
22

23. Main AEX membranes - Performances
Q resin Membrane 1 Membrane 2 Membrane 3 QyuSpeed D
Configuration Beads Rolled Sheet Open Pleated Stacked Flat Sheet Hollow fibers
Medium Material Agarose Cellulose PES PE PE
-N+-(CH3)3 -N+-(CH3)3 Quaternary -N+H-
Ligand Primary amine
amine (CH2CH3)2
Max Flow rate 1 CV/min 30 MV/min 10 MV/min 12.5 MV/min 13.5 MV/min
Pore Size - > 3 µm 0.8 µm 0.65 µm 0.2-0.3 µm
10% BSA DBC
> 100 > 29 > 60 > 50 > 40
(g/L-medium)
10% DNA DBC
<5 < 10 < 35 < 35 < 35
(g/L-medium)
at ~ 0 mS/cm
10% DNA DBC < 30
Not applicable.
<2 < 30 < 30
(g/L-medium) (if another Max 5 mS/cm
type)
at 15 mS/cm
Virus LRV > 3-4 > 3-4 > 3-4 >4 >4
Usually much lower reduction than for DNA and Viruses. Very much depending on the operating
HCP reduction
conditions and HCP characteristics (large pI range).
Number of
Up to 100 Single use Not precised Single use > 10
regenerations
23

24. Main AEX membranes - Performances
Q resin Membrane 1 Membrane 2 Membrane 3 QyuSpeed D
Configuration Beads Rolled Sheet Open Pleated Stacked Flat Sheet Hollow fibers
Medium Material Agarose Cellulose PES PE PE
-N+-(CH3)3 -N+-(CH3)3 Quaternary -N+H-
Ligand Primary amine
amine (CH2CH3)2
Max Flow rate 1 CV/min 30 MV/min 10 MV/min 12.5 MV/min 13.5 MV/min
Pore Size - > 3 µm 0.8 µm 0.65 µm 0.2-0.3 µm
10% BSA DBC
> 100 > 29 > 60 > 50 > 40
(g/L-medium)
10% DNA DBC
<5 < 10 < 35 < 35 < 35
(g/L-medium)
at ~ 0 mS/cm
10% DNA DBC < 30
Not applicable.
<2 < 30 < 30
(g/L-medium) (if another Max 5 mS/cm
type)
at 15 mS/cm
Virus LRV > 3-4 > 3-4 > 3-4 >4 >4
Usually much lower reduction than for DNA and Viruses. Very much depending on the operating
HCP reduction
conditions and HCP characteristics (large pI range).
Number of
Up to 100 Single use Not precised Single use > 10
regenerations
24

25. Main AEX membranes - Performances
Q resin Membrane 1 Membrane 2 Membrane 3 QyuSpeed D
Configuration Beads Rolled Sheet Open Pleated Stacked Flat Sheet Hollow fibers
Medium Material Agarose Cellulose PES PE PE
-N+-(CH3)3 -N+-(CH3)3 Quaternary -N+H-
Ligand Primary amine
amine (CH2CH3)2
Max Flow rate 1 CV/min 30 MV/min 10 MV/min 12.5 MV/min 13.5 MV/min
Pore Size - > 3 µm 0.8 µm 0.65 µm 0.2-0.3 µm
10% BSA DBC
> 100 > 29 > 60 > 50 > 40
(g/L-medium)
10% DNA DBC
<5 < 10 < 35 < 35 < 35
(g/L-medium)
at ~ 0 mS/cm
10% DNA DBC < 30
Not applicable.
<2 < 30 < 30
(g/L-medium) (if another Max 5 mS/cm
type)
at 15 mS/cm
Virus LRV > 3-4 > 3-4 > 3-4 >4 >4
Usually much lower reduction than for DNA and Viruses. Very much depending on the operating
HCP reduction
conditions and HCP characteristics (large pI range).
> 10
Number of
Up to 100 Single use Not precised Single use
regenerations (up to 100
possible)
25

26. QyuSpeed D advantages
 Shorter processing time.
 High salt tolerance.
 MV << CV (= less chromatographic medium).
 Lower buffer consumptions.
26

27. QyuSpeed D advantages
 Shorter processing time.
 High salt tolerance.
 MV << CV (= less chromatographic medium).
 Lower buffer consumptions.
 No packing.
 Less labor costs.
 Less hardware equipment.
 Smaller footprint.
 Easy/reliable scale up by adding modules.
27

28. QyuSpeed D advantages
QyuSpeed D
550 mL MV
Chromatography
column 50 L CV 28

29. PART 5 – Cost simulation:
Q resin, Single Use Q membrane
and QyuSpeed D,
for different batch sizes and
different numbers of batches/year

30. Cost simulation - Hypothesis
 Comparisons between QyuSpeed D, single use Q membrane, Q resin.
 MAb production from CHO cell culture with standard DSP steps.
 Each technology leads to similar purification yields.
 Protein concentration before AEX: 2 g/L.
 Conductivity of the MAb solution before loading: < 5 mS/cm.
 3 loading volumes:100 L (small), 1 500 L (medium), 5 000 L (large)
 Number of batches : 10/year, 40/year
30

31. Cost simulation - Hypothesis
 Comparisons between QyuSpeed D, single use Q membrane, Q resin.
 MAb production from CHO cell culture with standard DSP steps.
 Each technology leads to similar purification yields.
 Protein concentration before AEX: 2 g/L.
 Conductivity of the MAb solution before loading: < 5 mS/cm.
 3 loading volumes:100 L (small), 1 500 L (medium), 5 000 L (large)
 Number of batches : 10/year, 40/year
 Cost calculations based on 5 years operation.
 Q resin: 100 uses; QyuSpeed D: 15 uses.
 Buffer volume for packing step is not counted.
 P r i c e s :
- Q resin: 700 $/L-medium;
- Q membrane: 7 000 $/L-
medium; - QyuSpeed D: 14 000 $/L-
medium.
 Labor cost: 350 $/h (2 operators) 31

32. Buffer volumes for 1 cycle
Q membrane QyuSpeed D
Process step Q resin
(single use) (reusable)
Flush WFI
(CV* or MV**)
Pre-wash with buffer
(CV or MV)
Equilibration with buffer
(CV or MV)
Washing with buffer
(CV or MV)
Regeneration
with NaCl and NaOH
(CV or MV)
Total buffer volume per cycle
(CV or MV)
*CV: Column Volume
**MV: Membrane Volume
32

33. Buffer volumes for 1 cycle
Q membrane QyuSpeed D
Process step Q resin
(single use) (reusable)
Flush WFI
5 10 -
(CV* or MV**)
Pre-wash with buffer
5 10 10
(CV or MV)
Equilibration with buffer
5 10 15
(CV or MV)
Washing with buffer
5 10 10
(CV or MV)
Regeneration
with NaCl and NaOH 5 - 50
(CV or MV)
Total buffer volume per cycle
(CV or MV) 25 40 85
*CV: Column Volume
**MV: Membrane Volume
33

34. Buffer volumes for 1 cycle
Q membrane QyuSpeed D
Process step Q resin
(single use) (reusable)
Flush WFI
5 10 -
(CV* or MV**)
Pre-wash with buffer
5 10 10
(CV or MV)
Equilibration with buffer
5 10 15
(CV or MV)
Washing with buffer
5 10 10
(CV or MV)
Regeneration
with NaCl and NaOH 5 - 50
(CV or MV)
Total buffer volume per cycle
(CV or MV) 25 40 85
*CV: Column Volume Buffer cost: ~ 2 $/L
**MV: Membrane Volume
34

35. Costs for 1 cycle
Q membrane QyuSpeedD
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Loading volume (L) 100 1500 5000 100 1000 10000 100 1000 10000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane Volume (L)
Resin or Membrane cost (k$)
Buffer volume (L)
Buffer cost (k$)
Labor time (h)
Labor cost (k$)
Hardware cost (k$)
Validation cost (k$)
35

36. Costs for 1 cycle
Q membrane QyuSpeedD
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane Volume (L) 1 20 67 0,10 1 5 0,10 2 5
Resin or Membrane cost (k$)
Buffer volume (L)
Buffer cost (k$)
Labor time (h)
Labor cost (k$)
Hardware cost (k$)
Validation cost (k$)
36

37. Costs for 1 cycle
Q membrane QyuSpeedD
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane Volume (L) 1 20 67 0,10 1 5 0,10 2 5
Resin or Membrane cost (k$) 1 9 93 1 7 70 1 14 140
Buffer volume (L)
Buffer cost (k$)
Labor time (h)
Labor cost (k$)
Hardware cost (k$)
Validation cost (k$)
37

38. Costs for 1 cycle
Q membrane QyuSpeedD
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane Volume (L) 1 20 67 0,10 1 5 0,10 2 5
Resin or Membrane cost (k$)
Buffer volume (L) 33 500 1667 4 52 200 9 128 425
Buffer cost (k$)
Labor time (h)
Labor cost (k$)
Hardware cost (k$)
Validation cost (k$)
38

39. Costs for 1 cycle
Q membrane QyuSpeedD
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane Volume (L) 1 20 67 0,10 1 5 0,10 2 5
Resin or Membrane cost (k$)
Buffer volume (L) 33 500 1667 4 52 200 9 128 425
Buffer cost (k$) 0,07 1,00 3,33 0,01 0,10 0,40 0,02 0,26 0,85
Labor time (h)
Labor cost (k$)
Hardware cost (k$)
Validation cost (k$)
39

40. Costs for 1 cycle
Q membrane QyuSpeedD
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane Volume (L)
Resin or Membrane cost (k$)
Buffer volume (L)
Buffer cost (k$)
Labor time (h) 5 5 5 1,5 1,5 1,5 2,5 2,5 2,5
Labor cost (k$)
Hardware cost (k$)
Validation cost (k$)
40

41. Costs for 1 cycle
Q membrane QyuSpeedD
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane Volume (L)
Resin or Membrane cost (k$)
Buffer volume (L)
Buffer cost (k$)
Labor time (h) 5 5 5 1,5 1,5 1,5 2,5 2,5 2,5
Labor cost (k$) 1,8 1,8 1,8 0,5 0,5 0,5 0,9 0,9 0,9
Hardware cost (k$)
Validation cost (k$)
41

42. Costs for 1 cycle
Q membrane QyuSpeedD
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane Volume (L)
Resin or Membrane cost (k$)
Buffer volume (L)
Buffer cost (k$)
Labor time (h)
Labor cost (k$)
Hardware cost (k$) 140 180 350 - - - - - -
Validation cost (k$) 500 700 700 - - - 250 350 350
42

43. Costs for 1 cycle
Q membrane QyuSpeedD
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane Volume (L) 1 20 67 0,10 1 5 0,10 2 5
Resin or Membrane cost (k$) 1 9 93 1 7 70 1 14 140
Buffer volume (L) 33 500 1667 4 52 200 9 128 425
Buffer cost (k$) 0,07 1,00 3,33 0,01 0,10 0,40 0,02 0,26 0,85
Labor time (h) 5 5 5 1,5 1,5 1,5 2,5 2,5 2,5
Labor cost (k$) 1,8 1,8 1,8 0,5 0,5 0,5 0,9 0,9 0,9
Hardware cost (k$) 140 180 350 - - - - - -
Validation cost (k$) 500 700 700 - - - 250 350 350
43

44. First Comments
 Membrane chromatography technology leads to a much
lower volume of chromatography medium.
 The buffer consumptions are considerably reduced with
membrane chromatography.
44

45. Cost simulation
for 10 and 40 batches/year during 5 years
Q membrane QyuSpeed D
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Number of use 100 1 15
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Total cost for
10 batches/year
during 5 years (k$)
Total cost for
40 batches/year
during 5 years (k$)
45

46. Cost simulation
for 10 and 40 batches/year during 5 years
Q membrane QyuSpeed D
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Number of use 100 1 15
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Total cost for
10 batches/year 731 62 299
during 5 years (k$)
Total cost for
40 batches/year 1 005 247 447
during 5 years (k$)
46

47. Cost simulation
for 10 and 40 batches/year during 5 years
Q membrane QyuSpeed D
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Number of use 100 1 15
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Total cost for
10 batches/year 1 025 486 477
during 5 years (k$)
Total cost for
40 batches/year 1 458 1 946 856
during 5 years (k$)
47

48. Cost simulation
for 10 and 40 batches/year during 5 years
Q membrane QyuSpeed D
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Number of use 100 1 15
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Total cost for
10 batches/year 1 328 1 796 670
during 5 years (k$)
Total cost for
40 batches/year 2 160 7 185 1 628
during 5 years (k$)
48

49. Cost simulation
for 10 and 40 batches/year during 5 years
Q membrane QyuSpeed D
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Number of use 100 1 15
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Total cost for
10 batches/year 731 1 025 1 328 62 486 1 796 299 477 670
during 5 years (k$)
Total cost for
40 batches/year 1 005 1 458 2 160 247 1 946 7 185 447 856 1 628
during 5 years (k$)
49

50. Other Comments
 Membrane chromatography technology leads to a much
lower volume of chromatography medium.
 The buffer consumptions are considerably reduced with
membrane chromatography.
 Single use membrane chromatography is attractive with
small to medium loading volumes, when small number of
batches/year.
 QyuSpeed D is attractive with medium to large loading
volumes, whatever small or high number of batches/year.
 QyuSpeed D is highly cost effective thanks to its
regeneration capability.
50

51. Conclusion
 QyuSpeed D can replace current AEX
chromatography steps in your DSP.
 Very efficient removal of DNA, Viruses,
HCP, even at high conductivities.
 QyuSpeed D is a highly cost effective
technology thanks to its regeneration
capability.
51

52. ご清聴、ありがとうございま
す。

53. Backup slides

54. DNA DBC vs. conductivity
Internal study:
 Load solution: DNA 0.1g/L in 20mM Tris-HCl pH8.0, NaCl: 0M∼1.2M
 Flow rate：QSD: 2mL/min (3.6MV/min), Membrane A: 2mL/min (4.9MV/min)
40
0.3M NaCl
35
0.6M NaCl
10% DBC mg/mL-ad
30
25
Very high salt
20 QyuSpeed
tolerance for
15
Membrane A DNA DBC
10
0.9 M NaCl
5
1.2M NaCl
0
0 20 40 60 80 100
Conductivity mS/cm
54

55. Parvovirus removal
Internal study:
 0.5 vol% Serum Free-PPV spike (106/mL) + 10 g/L human-IgG in 0.1M NaCl
 Conductivity: 10 mS/cm, pH 7.9 (adjusted with NaOH)
 LRV evaluation: fraction 150 mL, Hemagglutination assay TCID50 method
 Flow rate: 5.0 MV/min
6.0
1250 MV, 1200 L/m2
4.5 PPV LRV > 4
for 16 kg IgG/L-ad
PPV LRV
3.0
(25 kg IgG/m2)
1.5
QyuSpeed D
0
0 750 1500 2250 3000
MV (mL sol./Membrane volume)
Throughput
55

56. IgG recovery and HCP removal
Internal study:
 10 g/L human-IgG + 0.1 mg/mL HCP
 0.15M NaCl; 20 mM Tris-HCl; pH 7.5
 Flow rate: 5.0 MV/min
100  Very high IgG recovery
at high loading volumes
90
80
 > 90% HCP removal
IgG Recovery ％
600 g IgG /L-media  Max IgG loading
70 depending on the
60 amount of impurities to
50
bind.
40 QyuSpeed D  Estimation: > 2000 g
IgG/L-medium
30
20
10
0
0 20 40 60 80 100
Loading volume (MV)
56

57. Process configuration
QyuSpeed D
Scale up
Waste
Product
Feed Equilibration Elution
/ CIP
57

58. QyuSpeed D operation steps
STEP- 1 Filter Connection
and Air Removal
Pre-washing
STEP- 2
and Leakage Test
STEP- 3 Membrane Equilibration
STEP- 4 Sample Loading
STEP- 5 Elution and Regeneration
58

59. Total Costs for 10 batches/Year during 5 years
Q membrane QyuSpeed D
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Number of use 100 1 15
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane cost (k$) 0,5 7 23 35 455 1750 4,67 70 233,3
Buffer cost (k$) 3,33 50 167 0,4 5 20 1 13 43
Labor cost (k$) 88 88 88 26 26 26 44 44 44
Validation cost (k$) 500 700 700 - - - 250 350 350
Hardware cost (k$) 140 180 350 - - - - - -
Total cost (k$) 731 1 025 1 328 62 486 1 796 299 477 670
59

60. Total Costs for 40 batches/Year during 5 years
Q membrane QyuSpeed D
Q resin
(single use) (reusable)
Loading capacity
150 2000 2000
(g protein/L-medium)
Number of use 100 1 15
Loading volume (L) 100 1500 5000 100 1500 5000 100 1500 5000
Protein loading (kg) 0,2 3 10 0,2 3 10 0,2 3 10
Resin or Membrane cost (k$) 1,9 28 93 140 1820 7000 18,7 280 933,3
Buffer cost (k$) 13,3 200 667 1,6 21 80 3 51 170
Labor cost (k$) 350 350 350 105 105 105 175 175 175
Validation cost (k$) 500 700 700 - - - 250 350 350
Hardware cost (k$) 140 180 350 - - - - - -
Total cost (k$) 1 005 1 458 2 160 247 1 946 7 185 447 856 1 628
60

61. ご清聴、ありがとうございま
す。