Facts On Quality And Safety Characteristics Of Fresh Frozen Plasma Single Donations And octaplasLG...
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Transcript of Facts On Quality And Safety Characteristics Of Fresh Frozen Plasma Single Donations And octaplasLG...
Facts On Quality And Safety Characteristics Of Fresh Frozen Plasma Single DonationsAnd octaplasLG®
Jürgen Römisch, Ph.D.
Senior Vice President R&D Plasma
Octapharma PPGmbH
Vienna, Austria
BPAC Meeting, Rockville, MDSeptember 20, 2012
2
Table of Contents
Quality and Safety Aspects of Fresh Frozen Plasma Single Donations
Plasma Transfusion Related Adverse Events (TRALI): Biochemical / cellular background
octaplasLG® - Manufacturing Process
octaplasLG® - Pathogen Safety
octaplasLG® - Plasma Quality / Biochemical Profile
3
Quality and Safety Aspects of FFP Single DonationsDonor screening requirements in USA for fresh frozen plasma
Unique donor identification and registration in a validated computer system
Deferral check (history of donor)
Donor questionnaire (preferably electronic); completed prior to each donation[1]
Education on blood borne diseases and transmission risks
Physical donor assessment by a health professional
Exclusion of donors who do not meet the established acceptance criteria (defined in 21 CFR 640.3 and other guidance documents)[2, 3]
Individual donations are tested on several parameters such as on the absence of HBsAg and antibodies against HIV, HCV and Syphilis
[1] Guidance for Industry: Implementation of acceptable full-length donor history questionnaire and accompanying materials for use in screening donors of blood and blood components.FDA, October 2006[2] http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfCFR/CFRSearch.cfm?fr=640.63[3] http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfCFR/CFRSearch.cfm?fr=640.3
4
Transfusion Related Acute Lung Injury has emerged as the leading cause of fatalities after blood/plasma transfusion
Incidence (TRALI Study Group)[4]: 2.57-0.81/10,000 transfused units (2006-2009)
Typical presentation of TRALI is development of dyspnea, hypoxemia, hypotension and fever 6 hrs after transfusion (bilateral pulmonary infiltrates)[5]
– Immune-mediated TRALI is usually caused by antibodies of the blood/plasma donor transferred to the recipient
Reactive antibodies to Human Leukocyte Antigens (HLA) or Human Neutrophil Antigens (HNA) cause complement and cell activation and aggregation
Release of cytokines and induction of pro-inflammatory reactions
Damage of vascular endothelium and exudation of fluid across the pulmonary basement membrane
Edema and associated/subsequent events
TRALI
Plasma Transfusion Related Adverse Events (TRALI) Biochemical/cellular background
[4] Transfusion Related Acute Lung Injury Letter; October 19, 2001http://www.fda.gov/BiologicsBloodVaccines/SafetyAvailability/ucm105850.htm[5] Toy P et al. For the TRALI Study Group. Transfusion-related lung injury: incidence and risk factors. Blood 2012; 119:1757-1767
5
octaplasLG® Manufacturing ProcessOverview: FFP pooling
Pooling of individual donations (630-1,520 FFP units) facilitates
Dilution and immune neutralization of - allergens - pathogenic antibodies - infectious agents
Leveling out individual heterogeneity, thus reducing variability of contents of plasma componentsresulting in a consistent product quality
YY
Y
Y Y
Y
Individual antibody specificities of different donations
Individual antigen structures contained in single FFP units
YYYYY
Cell fragments
Antibodies to WBCs were detected in a certain number of FFPs, but not in Octaplas® batches tested [6a,6b,7]
[6a] Sachs U. et al. Transfusion 2005; 45:1628-1631[6b] Barz D. et al. Anaesthesiol. Reanimate 1994; 156: 155-158[7] Sinnott P. et al. Eur. J. Immunogen. 2004; 31:271-274
6
octaplasLG® Manufacturing ProcessOverview: safety measures: pathogen safety
Antibodies to WBCs (TRALI)Non-Enveloped Viruses ADRs related to cells and fragments
Lipid-Enveloped Viruses
Bacteria & Parasites
Safety against
Infectious Prions
Fast thawing and pooling of single FFP units
Removal of cells, fragments and aggregates by 1.0 µm filtration
Full quality control and release of final product
Castor oil extraction of TNBP, clear filtration and solid phase extraction of Octoxynol
Addition of glycine
Affinity chromatography for PrPsc capture
Sterile filtration (0.45 µm and 0.2 µm)
Aseptic filling, sealing of bags and fast freezing (≤ -60°C) and storage (≤ -30°C)
S/D treatment (1% TNBP & 1% Octoxynol for 1-1.5 hrs at +30°C ± 1°C)
Pooling of individual donations (630-1,520 FFP units) facilitates
Dilution and immune neutralization of - allergens - pathogenic antibodies - infectious agents
Leveling out of individual heterogeneity and reducing variability of contents of plasma components, resulting in a consistent product quality
7
Pathogen SafetySafety measures
Beyond the basis precautionary safety measures ensured for FFP, such as donor screening and look-back procedures, the following octaplasLG® process steps add to the pathogen safety profile
– Plasma pool testing (HIV, HBsAG, HAV, HBV, HCV, HEV and Parvovirus B19)
– Solvent / Detergent (S/D) treatment for the inactivation of the most hazardous transfusion related enveloped viruses such as HIV, HBV, HCV and WNV– and other potentially emerging lipid-enveloped viruses
– Immune neutralization by specific antibodies in the plasma pool and the final product such as HAV, Parvovirus B19 and HEV– Pooling of on average 1,000 plasma donations ensures the presence of those antibodies– Respective minimum antibody titers are specified and must be met for product release
– Micro-filtration minimizes the risk of bacteria and parasites presence
– Specific affinity chromatography with a high binding capacity can remove potentially present infectious prion protein, which is not inactivated or sufficiently removed by other manufacturing steps
8
Plasma pool testing
Pathogen Safety
Parameter* Limit Plasma Pool*
anti-HIV 1/2 negative
HBsAg negative
anti-HAV** ≥ 1 IU/ml
HAV-PCR negative
HBV-PCR negative
HCV-PCR negative
HEV-PCR# negative
HIV-PCR negative
Parvo B19 PCR ≤ 10.0 IU / µl
The plasma pool for octaplasLG® (630-1,520 FFP units) is tested for:
* under FDA review ** tested in IP sample 1 # Implementation Nov. 2012
9
Pathogen SafetyS/D treatment
Solvent / Detergent (S/D) treatment causes disruption of the lipid envelopes of LEV resulting in complete, fast and robust virus inactivation[8,9]
PRV: model virus for HBV; BVDV: model virus for HCV
[8] Dichtelmüller H.O. et al. Transfusion 2009; 49:1931-1943[9] Hellstern P. et al. Transfus. Med. Hemother. 2011: 38:65-70
Red line — = kinetics – Blue square = below limit of detection (LOD)
0 10 20 30 40 50 600
1
2
3
4
5
6
7
8
9
WNV
Time [Minutes]
Mea
n V
iru
s L
oad
[lo
g10
TC
ID50
]
0 10 20 30 40 50 600
1
2
3
4
5
6
7
8
9BVDV
Time [Minutes]
Mea
n V
iru
s L
oad
[lo
g10
TC
ID50
]
0 10 20 30 40 50 600
1
2
3
4
5
6
7
8
9PRV
Time [Minutes]
Mea
n V
iru
s L
oad
[lo
g10
TC
ID50
]
≥ 5.09 log10
≥ 5.24 log10 ≥ 5.99 log10
At 85% S/D
concentration
(robustness)
0 10 20 30 40 50 600
1
2
3
4
5
6
7
8
9
HIV
Time [Minutes]
Mea
n V
iru
s L
oad
[lo
g10
TC
ID50
]
≥ 5.47 log10
At 85% S/D
concentration
(robustness)
Total inactivation 4 log10 to LOD within 1 minute, i.e. 98.9% time-safety-margin for 1 hours S/D
10
Pathogen SafetyImmune neutralization
Immune neutralization depends on the virus load and the specific antibody content of the plasma pool and the final container
For non-enveloped viruses, which are not inactivated by S/D treatment, minimum titers of neutralising antibodies were specified, thus must be present
Virus Plasma pool octaplasLG®
HAV PCRanti-HAV IgG
negative≥ 1 IU/ml ≥ 1 IU/ml
B19 PCRanti-B19 IgG
≤ 10.0 IU/µl≥ 11 IU/ml
HEV PCRanti-HEV IgG
Implementation Nov. 2012≥ 0.2 IU/ml
HAV: Hepatitis A VirusB19: Parvovirus B19HEV: Hepatitis E Virus
11
Pathogen SafetyMicro-Filtration
Filtration steps at different stages of the manufacturing process minimize the risk of a bacterial or parasite transfection
– 1.0 µm filtration subsequent to FFP pooling– 0.45 µm and 0.2 µm sterile filtrations prior to aseptic filling
12
Pathogen SafetyGlobal virus reduction factors during octaplasLG® manufacturing
OctaplasLG. Biological Licence Application (STN 125416/0). 3.2.A.2.4.3 Summary Tables of Virus Validation Factors.
PCR of HAV, B19, HBV, HCV, and HIV genomes are performed on plasma pool level; HEV PCR is being implemented
Anti- HAV (pool and product), HBsAg (pool), anti HIV 1/2 (pool), anti-B19 (product) and anti-HEV (product) antibody titers are tested and must comply with the defined specifications
Step HIV-1 PRV SBV BVDV WNV VACV HSV-1 HAV COX-B6 POL-1
Immune neutralisation (log10)
n.a. n.a n.a n.a n.a n.a≥ 11.1(pool)
≥ 10.0(pool)
≥ 8.6(pool)
≥ 10.9(pool)
S/D treatment (log10)
≥ 6.18 ≥ 5.03 ≥ 5.31 ≥ 5.12 ≥ 5.63 ≥ 5.00 n.a n.a n.a n.a
Global reduction factor (log10)
≥ 6.18 ≥ 5.03 ≥ 5.31 ≥ 5.12 ≥ 5.63 ≥ 5.00≥ 11.1(pool)
≥ 10.0(pool)
≥ 8.6(pool)
≥ 10.9(pool)
Lipid-enveloped viruses Non-enveloped viruses
HIV: Human Immunodeficiency Virus Type 1 PRV: Pseudorabies Virus; model for Herpes VirusSBV: Sindbis Virus; model for Hepatitis C Virus BVDV: Bovine Viral Diarrhea Virus; model for Hepatitis C VirusWNV: West Nile Virus VACV: Vaccinia VirusHSV-1. Herpes Simplex Virus Type 1 HAV: Hepatitis A VirusCOX-B6: Coxsackie Virus B6 POL-1: Poliovirus type 1
13
Pathogen SafetyPrPSc Affinity Chromatography
A sensitive and robust assay to detect eventually present infectious prion protein in blood and plasma donations is not yet available
The affinity chromatography with a specific immobilized ligand has a high capacity to adsorb infectious prion protein, thus reduces the risk of transfusion related Creutzfeldt-Jakob Disease
– Prion removal capacity (PrPSc) of octaplasLG® was confirmed in animal bioassay studies[10]
– 5.64 log10 ID50 / ml gel
[10] Heger A. et al. Vox Sang 2012; 104:294-301.
14
Plasma Quality & Biochemical ProfileQC release parameters
For product release each octaplasLG® batch is tested on a number of parameters (following slides) in the Quality Control department
Specifications for these parameters have been set and some of them were revised for US for octaplasLG®, also taking into consideration the history in US of a first generation S/D treated plasma product from another manufacturer[11]
These specifications must be met for release of each batch to safeguard the presence of physiologically relevant concentrations of the tested plasma components, in particular
– Balance of coagulation factors and their haemostasis regulating proteins (inhibitors)
[11] Coignard P. et al. Hepatology 2002; 36 (4): Pt.2BPAC 102nd Meeting May, 2012: Evaluation of potential new plasma products manufacturedfollowing storage at room temparature for up to 24 hours
15
Plasma Quality & Biochemical ProfileProtein S and Plasmin Inhibitor
In the course of the implementation of the Ligand Gel chromatography the S/D treatment exposure time was shortened
– maintaining a sufficient safety margin for complete inactivation of lipid-enveloped viruses
– but facilitating an improved plasmin inhibitor in-vitro recovery[12,13]
[13]
[12] Heger A. et al. Vox Sang 2012; 103:30[13] Heger A. et al. Vox. Sang. 2009; 96:225
16
Plasma Quality & Biochemical ProfileProtease inhibitors and cofactors
ParametersFFP
(n=12)[14]
Final Product Specification
octaplasLG® USA #
octaplasLG®
(n=33)[12]
Antithrombin [IU/ml] 1.06 (0.77-1.23) n.s. 0.92 (0.74-1.00)
Heparin cofactor II [IU/ml] 0.95 (0.76-1.19) n.s. 1.06 (0.82-1.44)
Protein C [IU/ml] 0.89 (0.79-1.05) ≥ 0.7 0.97 (0.79-1.10)
Protein S [IU/ml] 1.03 (0.71-1.39) ≥ 0.4 0.61 (0.50-0.72)
Plasmin inhibitor [IU/ml] 1.04 (0.95-1.18) ≥ 0.4 0.52 (0.30-0.64)
α1-Antitrypsin [mg/ml]* 1.57 (0.98-2.22) n.s. 1.29 (0.88-1.76)
C1-inhibitor [IU/ml] 0.87 (0.72-0.97) n.s. 0.89 (0.67-1.23)
n.s., not specified
*different test methods used for FFP (kinetic nephelometry) and octaplasLG® (ELISA)
# under review by FDA
[12] Heger A. et al. Vox. Sang. 2012; 103:130[14] Beeck H. et al. Vox. Sang. 1998; 74:219-223
17
Plasma Quality & Biochemical ProfileCoagulation factors and proteins of the haemostatic system
ParametersFFP
(n=12) [14]
Final Product Specification
octaplasLG® USA
octaplasLG®
(n=33) [12]
Fibrinogen [mg/ml] 2.6 (1.9-3.6) 2.0-4.0 2.7 (2.5-3.1)
Factor II [IU/ml] 0.88 (0.77-1.18) ≥ 0.7 0.92 (0.79-1.08)
Factor V [IU/ml] 0.90 (0.73-1.50) ≥ 0.6 0.81 (0.70-1.00)
Factor VII [IU/ml] 0.95 (0.67-1.38) ≥ 0.6 1.03 (0.70-1.20)
Factor VIII [IU/ml] 0.76 (0.52-1.13) ≥ 0.5 0.85 (0.60-1.30)
Factor IX [IU/ml] 1.02 (0.82-1.28) n.s. 0.94 (0.74-1.30)
Factor X [IU/ml] 0.79 (0.62-0.99) ≥ 0.8 0.93 (0.85-1.04)
Factor XI [IU/ml] 1.13 (0.96-1.80) ≥ 0.7 0.88 (0.80-1.00)
Factor XII [IU/ml] 0.82 (0.45-1.12) n.s. 1.00 (0.81-1.35)
Factor XIII [IU/ml] 1.06 (0.68-1.69) n.s. 1.01 (0.85-1.20)
VWF:RCo [IU/ml] 0.89 (0.77-0.98) n.s. 0.88 (0.72-1.01)
ADAMTS13 [IU/ml][15] 1.24 (1.07-1.44) ≥ 0.7 0.91 (0.75-1.30)
Plasminogen [IU/ml] 0.98 (0.82-1.39) n.s. 0.91 (0.75-1.03)
n.s.: not specified[12] Heger A. et al. Vox. Sang. 2012; 103:130[14] Beeck H. et al. Vox. Sang.1998; 74:219-223[15] Heger A. et al. Vox. Sang. 2007; 92:206-212 (Octaplas, n=18)
18
Plasma Quality & Biochemical ProfileRanges of plasma protein concentrations in octaplasLG® batches and FFPs are equivalent
Facto
r II
Facto
r VII
Facto
r IX
Facto
r XI
Facto
r XIII
ADAMTS13
HCII
Prote
in S
A1-AT
Facto
r V
Facto
r VIII
Facto
r X
Facto
r XII
VWF:R
co
ATIII
Prote
in C
Plasm
in in
h
C1-IN
H
0
20
40
60
80
100
120
140
160
180
octaplasLG® batches (n=33) Single-donor FFP bags (n=12)
Po
ten
cy [
%]
[12] Heger A. et al. Vox. Sang. 2012; 103:130[14] Beeck H. et al. Vox. Sang. 1998; 74:219-223
19
The Thrombin Generation Assay demonstrated the overall coagulation potential of octaplasLG®, comparable to the range measured in FFPs
Plasma QualityGlobal coagulation assays and specific assays
[12] Heger A. et al. Vox Sang. 2012; 103:130[16] Pock K. et al..Transfusion Apher. Sci, 2007; 37:223-231
OctaplasLG® (US plasma, n=12)
[12,16]
ParametersFFP
(n=18) [16]
octaplasLG®
(n=33) [12]
Peak thrombin [nM] 331 (195-437) 396 (286-507)
Lag phase [min] 14.6 (5.3-20.5) 12.2 (10.4-16.0)
20
ADAMTS13 levels and von Willebrand Factor multimeric structures are equivalent in octaplas® batches and in FFP [15,17]
Plasma QualityGlobal coagulation assays and specific assays
1 2 3 4 5 6 7 8 9 10 11
HighVWF-MM
IntermediateVWF-MM
LowVWF-MM
HighVWF-MM
IntermediateVWF-MM
LowVWF-MM
octaplas®
Normal Plasma Ref. Standard
ADAMTS13
VWF
VWF multimer structure: densitometry FFP octaplas®
[15] Heger A. et al. Vox Sang. 2007; 92:206-212 [17] Heger A. et al. Haemophilia. 2006; 12 (2):05PO125
21
Summary
In addition to the basic safeguarding measures for FFP with respect to a potential transfusion related infection, the octaplasLG® manufacturing process adds S/D treatment, micro-filtrations and a dedicated PrPSc adsorption step (LG); immune neutralization of non-enveloped viruses and PCR testing in the plasma pool are ensured for each octaplasLG® batch
FFP pooling of (on average 1,000 donations) results in neutralization of pathogenic antibodies and substances (minimization TRALI risk)
The overall biochemical profiles of FFPs and octaplasLG® are comparable
– Reduction of S/D exposure period increased the plasmin inhibitor in-vitro recovery in octaplasLG®
– Balanced coagulation factor and inhibitor contents are ensured by product release specifications (several inhibitor activities) for each octaplasLG® batch
22
Acknowledgements
Andrea Heger
Simone Meindl
Andrea Neisser-Svae
Barbara Rangetiner
Torben Schmidt
Tor-Einar Svae
Michael Szkutta