Sepacell Prima TM Asahi Combination Filter, Sepacell Prima TM for Prion and Leukocyte Reduction...
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Transcript of Sepacell Prima TM Asahi Combination Filter, Sepacell Prima TM for Prion and Leukocyte Reduction...
Sepacell PrimaTM
Asahi Combination Filter, Asahi Combination Filter, Sepacell PrimaSepacell PrimaTMTM
for Prion and Leukocyte Reductionfor Prion and Leukocyte Reduction
Asahi Kasei MedicalTomo Yokomizo, Morikazu Miura
29th October, 2010
Sepacell PrimaTM
Asahi Kasei MedicalAsahi Kasei MedicalOne of major suppliers of LR filter
Many types of LR filter, used all over the world
Nano-filtration technology for virus removal in plasma fractionation, etc.
Sepacell PrimaTM
Contents
Concept and Technology Development Status
Prion Reduction Western blot method Bioassay test
Leukocyte Reduction RCC Quality Biological Safety
Next Step
- Sepacell PrimaTM -
Sepacell PrimaTM
The Concept of Filter
Reducing prions from RCC Covering current LR filter functions
No Additional Operating Time No Impact on Customer SOP No Further RCC loss
One Step FilterOne Step FilterSimultaneous reduction of prions and Simultaneous reduction of prions and ll eukocyteseukocytes
Sepacell PrimaTM
Key Points of Technology
Materials Base fibers: Polyester Surface: New material
Prion reduction mechanism Adsorption: High affinity to prion
Filter design Equivalent to the current Red Cell fil
ter in Europe, Sepacell® Pure RC.
RCC
Filter
SAGM
Sepacell PrimaTM
Scheme of Prion Spiking Test
FilterFilter
Microsomal fraction (10w/v%)
Scrapie infected hamster with 263k scrapie strain
Brain homogenate
Leukoreduced RCC used to eliminate phagocytic effect
Western blot
LR SAG-M RCC
Sepacell PrimaTM
Prion Spiking Test Results (1)
Test laboratories X Y
Filter Pool 1 Pool 2 Pool 3 Pool 4
Log10
Reduction
Factor
A > 3.0 > 3.0 > 3.0 > 4.5
B > 3.0 > 3.0 > 3.0 -
C - - - > 4.5
D 0.5 0.2
A: Sepacell PrimaTM sterilized with SteamB: Sepacell PrimaTM sterilized with E-BeamC: Sepacell PrimaTM sterilized with Steam twiceD: Control (Pall WBF2)
•Prion reduction confirmed at two different test sites •Sterilization tolerance to both Steam and E-Beam
Vox Sanguinis 2008; 95 (suppl. 1): 271, P-588
Sepacell PrimaTM
Prion Spiking Test Results (2)
Filter RCC
Added
FFP
(mL)
Plasma
Protein
(mg/mL)
Log10
Reduction
Facotr
Control (Pure RC)
LR-RCC
0 3.2 0.92
Sepacell PrimaTM 0 3.2 > 4.5
Sepacell PrimaTM 9.5 7.2 > 4.5
Sepacell PrimaTM 22 10.8 > 4.5
Sepacell PrimaTM NLR-RCC - - > 4.5
•No impact observed within plasma protein variation of RCC•No negative effect induced by leukocytes observed
LR-RCC: Leukocyte reduced RCC, pooled and split before FFP additionNLR-RCC: Non - leukocyte reduced RCC
Transfusion 2008; 48 (suppl.): 86A, SP141
(at the laboratory Y)
Sepacell PrimaTM
Scheme of Bioassay Test
FilterFilter
Microsomal fraction (10w/v%)
Scrapie infected hamster with 263k scrapie strain
Brain homogenate
Leukoreduced RCC used to eliminate phagocytic effect
Dilution series from pre / post filtration samples
0.05 mL injected intracerebrally to Golden Syrian hamsters
Observed up to 365 days
LR SAG-M RCC
Sepacell PrimaTM
Bioassay Test Results
After 365 day observation
Log(ID50) reduction is 4.20 Log10
based on Spearman-Kärber method.-Log10ID50 = Log10 starting dilution – ([∑p – 0.5] x Log10 dilution factor)
•Prion reduction performance supported by Bioassay test
Vox Sanguinis 2009; 97 (suppl. 1): 101, P-092
No abnormal or unexpected observation in the groups of;•Positive control (263k microsomal post nuclear fraction)•Sham injected control (HBSS)•Uninjected control
100 10-1 10-2 10-3 10-4 10-5 10-6
mean 154 213 ( > 365 )
sd 34 65 -
0% 28% 100%
mean 127 145 168 ( > 365 )
sd 8 34 40 -
0% 0% 17% 100%
Dilution series
PostFiltration
PreFiltration
Survival Rate
Survival Rate
Survival Period(Days)
Survival Period(Days)
Sepacell PrimaTM
LR Filter Performance (1)
Residual Leukocytes ( Log WBC/unit)
Hemoglobin ( g/unit)
Ambient Temp. Hold, Day1 Filtration
* No significant difference
* No significant difference
Combination Filter PureRC
n=12 n=12
mean* 4.19 4.43
σ 0.32 0.34
Sepacell PrimaTM Control (Pure RC)
Combination Filter PureRC
n=12 n=12
mean* 49.2 48.5
σ 4.7 6.5
Sepacell PrimaTM Control (Pure RC)
Sepacell PrimaTM
LR Filter Performance (2)
Residual Leukocytes ( Log WBC/unit)
Hemoglobin ( g/unit)
4 Degree Hold, Day2 Filtration
* No significant difference
* No significant differenceTransfusion Medicine 2008; 18 (suppl.): 30, P08
Combination Filter PureRC
n=48 n=12
mean* 4.32 4.65
σ 0.60 0.56
Sepacell PrimaTM Control (Pure RC)
Combination Filter PureRC
n=48 n=12
mean* 50.6 51.4
σ 3.5 3.5
Sepacell PrimaTM Control (Pure RC)
Sepacell PrimaTM
RCC Quality
Parameters evaluated Hemolysis (%) C3a (ng/mL) Pottasium (mmol/L) pH Lactate (mg/dL) 2,3 – DPG (mmol/L) ATP (micro-mol/dL) Glucose (mg/dL)
Tested at day 0, 21 and 42 Sample (Sepacell PrimaTM, n=9) Control (Pure RC, n=3)
The results were comparable between sample and controlin all the test
Sepacell PrimaTM
Hemolysis results
Met CE guideline
Transfusion Medicine 2008; 18 (suppl.): 30, P08
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
Pre filtration Post filtration 21days 42days
Hem
olys
is(%
)
samplecontrol
Sepacell PrimaTM
Biological Safety Test Results
Test Item EB Steam
Cytotoxicity Pass
Sensitization Pass
Intracutaneous Reactivity Pass
Systemic Toxicity (acute) Pass
Haemolysis Testing Pass
Overall results show the sufficient safety of new filter.
Ames Test with New Polymer negative
ISO 10993
Vox Sanguinis 2008; 95 (suppl. 1): 271, P-588
Sepacell PrimaTM
Summary
* C3a, Pottasium, pH, Lactate, 2,3-DPG, ATP, Glucose
Items Requirement Present Status
WB Method: > 3 Log
Exogenous: > 4 Log
Residual Leukoctyes < 1x106/unit Same as PureRC
Hemoglobin min 40g/unit Same as PureRC
Hemolysis <0.8% at 42 days ca. 0.4% at 42 days
Other RBC Quality Factors* Same as existing LRF Same as PureRC
Sterilization Steam & Irradiation Both Applicable
ISO-10993 No issue found
Ames Test Negative
Prion Reduction
Biological Safety
> 3 Log
Sepacell PrimaTM
Next step
Sepacell PrimaTM to be CE marked in SCD kit Ready to provide samples for in-vitro test To be ready for clinical test soon
Asahi in-house tests are on-going Quality control method established
Sepacell PrimaTM
Prion Reduction Test
Spiking Study by Western Blot and Bioassay test Facility: Laboratories in Europe and US
Materials
Agent: The hamster 263K strain of scrapie
Spiking materials: Microsomal Fraction
Medium: Leukoreduced* SAGM-RCC
* to avoid the possibility of WBC related removal of PrPSC, such as phagocytic effect and non-specific binding.
Sepacell PrimaTM
RCC Quality Test Results
Hemolysis(%)
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
Pre filtration Post filtration 21days 42days
Hem
olys
is(%
)
samplecontrolmean, SD
C3a(ng/ mL)
0
500
1000
1500
2000
Pre filtration Post filtration 21days 42days
C3a
(ng/
mL)
samplecontrolmean, SD
Pottasium(mmol/ L)
0
5
10
15
20
25
30
35
40
45
Pre filtration Post filtration 21days 42days
Pot
tasium
(mm
ol/L
)
samplecontrolmean, SD
pH
6.4
6.6
6.8
7.0
7.2
7.4
Pre filtration Post filtration 21days 42days
pH
samplecontrolmean, SD
Lactate(mg/ dL)
0
50
100
150
200
250
300
350
Pre filtration Post filtration 21days 42days
Lact
ate(
mg/
dL)
samplecontrolmean, SD
2,3-DPG(mmol/ L)
0.0
0.5
1.0
1.5
2.0
2.5
Pre filtration Post filtration 21days 42days
2,3-
DPG
(mm
ol/L
)(%)
samplecontrolmean, SD
ATP(μ mol/ dL)
0
20
40
60
80
100
120
Pre filtration Post filtration 21days 42days
AT
P(μ
mol
/dL)
samplecontrolmean, SD
Glucose(mg/ dL)
0
200
400
600
800
1000
Pre filtration Post filtration 21days 42days
Glu
cose
(mg/
dL)
samplecontrolmean, SD
•Comparable RCC quality to control filter
Transfusion Medicine 2008; 18 (suppl.): 30, P08