Factors for Life.

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3

Content

1. CSL Behring – your partner in coagulation disorders ...................... 5

2. Factors for life .................................................................................... 8

3. Plasma derived products .................................................................. 11

4. Recombinant products ..................................................................... 24

5. Summary ........................................................................................... 28

6. Abbreviations ................................................................................... 28

7. Literature .......................................................................................... 30

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1. CSL Behring – your partner in coagulation disorders

Coagulation factor concentrates may be useful in a wide range of clinical situations from inborn de-fi ciencies to acute medical or sur-gical conditions.Globally more than 1% of the world‘s population is affected by bleeding disorders such as haemo-philia A, haemophilia B or von Willebrand disease (VWD). In se-vere cases their treatment invol-ves the prompt and proper use of clotting factor concentrates. Haemophilia A is caused by a de-fi ciency of clotting factor VIII and accounts for about 80 % of all haemophilia cases. Haemophilia B results from factor IX defi ciency. Nearly 1 % of the people world-wide have defects in the genetic coding for von Willebrand factor (VWF) leading to different types of VWD. Oral anti-coagulant therapy be-ars an increased bleeding risk even at therapeutic doses and an even more pronounced risk when overdosed. Due to a variety of interactions this is not unusual. Acute major bleeds or emergen-cy interventions require reversal of oral anticoagulation with a prothrombin complex concentra-te (PCC), attempting to avoid a haemostatic crisis.Critically ill patients may develop severe dyscoagulative conditions e.g. disseminated intravascular coagulation (DIC) or dilution co-agulopathy. These patients may benefi t from treatment with spe-cifi c factor concentrates replacing

the defi cient proteins necessary for a proper coagulation.

The goal of therapy is to decrease the frequency and severity of bleeds and to prevent the long-term complications. This is achie-ved by raising factor levels. Treat-ment can be administered either as on demand therapy where bleeds are treated upon occur-rence or as prophylaxis. The indi-vidually tailored therapy enables patients to have an improved quality of life. For non-haemo-philic patients in the intensive care setting therapy is aiming at avoiding imbalances in the co-agulative system leading to fatal bleeds or (micro)-thrombotic con-ditions.

Products available are safe, effec-tive and well investigated. CSL Behring is committed to provide one of the broadest ranges of factor concentrates for coagula-tion disorders worldwide.

Products are made by two well-established technologies, plasma-derived products purifi ed from natural plasma or concentrates manufactured by recombinant technology that utilises the inser-tion of human genetic material into a host cell of a foreign spe-cies.

From the donated plasma a range of therapeutic proteins are ex-tracted in a careful manner to re-

tain their biological integrity. This process provides opportunities to obtain a variety of products from this valuable raw material.

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Virus safety is one essential aspect of product quality and CSL Behring does everything possi-ble in order to reach the highest standards. Strict donor selection criteria have been implemented and a large number of tests are performed to minimise a poten-tial virus and prion load in the basic raw material. Methods for clearing viruses from plasma can be divided in two categories: In-activation processes (e.g. pasteu-risation) and removal steps (e.g. precipitation, absorption, virus fi ltration, monoclonal antibody purifi cation).

Pasteurisation is a unique proce-dure of pathogen inactivation and was introduced by CSL Behring (at that time called Behringwerke) in 1979. It is a very effective me-thod to inactivate enveloped and non-enveloped viruses is in our hands gentle enough to preserve the high quality of the products. Even though the HIV virus was not known at that time, it was later shown that pasteurisation effectively inactivated this virus in addition to other viruses.

Almost all steps of the manufac-turing process contribute to the safety of clotting factor concen-trates, and over the last 25 years the risk of virus transmission by the application of plasma derived products has become a theoreti-cal risk. CSL Behring’s long experience in providing safe and effective factor concentrates makes the company an ideal partner in the treatment of coagulation disor-ders and other situations requi-ring potentially life saving factor preparations.

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2. Factors for life

Figure 1: The coagulation cascade

Table 1: Product portfolio of CSL Behring

Many options for treatment of bleeding situations*

Products

FI FII, VII, IX, X FVIII FIX FX FXIII VWF ATIII

Haemo-complettan® P

Beriplex® P/N Beriate® PHelixate® NexGenHelixate® FSMonoclate®-PHaemate® PHumate-P®

Berinin® PMononine®

Factor X P Behring

Fibro-gammin® P

Haemate® PHumate-P®

Kybernin® P

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The coagulation of blood is a com-plex mechanism. Early models of the coagulation cascade were di-vided into two parts: the “intrin-sic” and the “extrinsic” pathway. But this was inconsistent with cli-nical observations in several key respects and did not fully explain the pathways leading to haemos-tasis in vivo. To address this a new model was developed.

It consists of four consecutive overlapping stages: initiation, amplifi cation, propagation and stabilisation and explains some aspects of haemostasis that a pro-tein-centric model does not.

Initiation: This process takes place on the surface of tissue factor (TF)-bearing cells, such as fi broblasts and platelets. Factor VII binds to cellular TF and is activated. This FVIIa/TF complex activates FX. FXa generates a small amount of thrombin which plays a role in the amplifi cation stage. Amplifi cation: Through the in-teraction of VWF, platelets ad-here to the intravascular lesion and become activated. The small amount of thrombin generated in the initiation phase activates factors V, VIII and XI. Assembly of the tenase- and prothrombinase- complexes and large scale throm-bin generation is initiated.

Propagation: The large scale thrombin generation catalyses the conversion of fi brinogen to fi brin and a clot is formed. This process leads to the activation of FXIII, which is necessary for the fi -nal stage.

Stabilisation: The covalent cross linking of the fi brin polymers that make the clot insoluble.

Indication Product

Haemophilia A

Beriate® PHelixate® NexGen/FSMonoclate®-PHaemate® P; Humate-P®

von Willebrand Disease Haemate® P; Humate-P®; Stimate®

Haemophilia BBerinin® PMononine®

Life-threatening haemorrhages with Vit-K dependent factor defi ciency Beriplex® P/N (Prothrombin Complex Concentrate)

Life-threatening haemorrhages with fi brinogen defi ciency Haemocomplettan® P

Defi ciency of Factor X (Stuart-Prower factor) Factor X P Behring

Factor XIII defi ciency Fibrogammin® P

Prophylaxis and therapy of thrombembolic episodes in AT III defi ciency Kybernin® P

* Not all products are licensed globally and approved indications may divert between countries.

Table 2: Products and indications

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3. Plasma derived products

Donor selection and testing

Criteria for donor selection

First time donors• Good health and a fl awless medical history • Check of vital signs, physical examination, lab analyses• Infection screen of donation• Quarantine until the next donation is screened negative

Regular donors• Good health and medical history update• Check of vital signs• Infection screen of donation• Minimum 60 days of inventory hold to cover

diagnostic window.

Exclusion of at-risk donors

Figure 2: Criteria for plasma donor selection

Product safety starts with the do-nor selection. Thus collection cen-tres for sourced plasma are moni-tored and typically located in areas of the U.S. or north/middle Europe with low incidence of HIV and he-patitis. Each candidate donor un-

dergoes multiple testing proce-dures, including complete physical examination, medical history and laboratory analyses. The fi rst do-nation from a sourced plasma donor undergoes a quarantine storage up to 6 months duration.

Only when the donor returns for a second donation within this time period and again is tested nega-tive, will the fi rst donation be released for further processing.

Table 3: Testing procedure at CSL Behring – voluntary industry standard is always met* For the majority of products

Rejection of positive donations

Pathogen Serological testing of every donation

Mandatory NAT/PCR testing in fractionation pool

Voluntary Industry standard NAT/PCR testing in plasma pool (PPTA)

NAT/PCR test performed in plasma pool by CSL Behring*

HIV Anti-HIV 1Anti-HIV 2

–

HCV (Hepatitis C) Anti-HCV

HBV (Hepatitis B) HBsAg –

HAV (Hepatitis A) – – –

Parvovirus B19 – –

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The laboratory analyses of each donation includes serological tests for HIV-1/2 antibodies, hepati-tis B virus (HBV) surface antigen (HBsAg) and hepatitis C Virus (HCV) antibodies. As the time from infection to the develop-ment of antibodies (the diagnostic window) varies between the dif-ferent viruses, there is a need for an additional test to minimise the risk of processing a donation contaminated with virus antigens or pathogenic viral genetic mate-rial. CSL Behring implemented a

sensitive nucleic acid amplifi ca-tion technique (NAT) known as polymerase chain reaction (PCR) to further reduce the potential virus load of plasma pools. To mi-nimise the exclusion of relevant raw material (plasma), a pooling strategy of minipools has been implemented allowing to discard NAT/PCR reactive minipools and/or donations only. CSL Behring‘s NAT/PCR test** will detect geno-mic sequences of HBV, HCV, HAV, HIV-1 and high titres of Parvovi-rus B19. In addition to this donor/

minipool-testing, CSL Behring per-forms an in process control on the manufacturing pools to increase safety.

Since introduction, the NAT/PCR testing strategy has been proven to be very effective in reducing the virus load in the plasma pool, resulting in a signifi cant increase of the safety margin.

Measures to minimize the risk of prion transmission

• No donors with a family history of CJD• No donors who received human pituitary hormones/dura mater or cornea transplantation• No plasma from the U.K. • Integration of prion reduction steps into manufacturing process

Until now prions could never be demonstrated in human plasma. However, four cases* of variant Creutzfeldt-Jakob-disease (vCJD, associated with mad cow disease or bovine spongiform encephe-lopathy, also known as BSE) have been confi rmed in the U.K. in recipients of non-leukodepleted red cells donated by donors, that a few years later were diagnosed with vCJD. Therefore the criteria for donor selection have been re-viewed and CSL Behring has imp-lemented these criteria:

Donors with a family history of CJD or who were treated with human pituitary hormones or received a dura mater or cornea transplantation are excluded. Ad-ditionally, there is no processing of plasma from the U.K. All gui-delines and regulations regarding collection of plasma and donor deferral are met wherever plasma is collected for CSL Behring or the fi nal products are distributed.

Furthermore CSL Behring has in-tegrated effective prion reduc-

tion steps into the manufacturing processes for a further decrease of the potential risk. For Haema-te® P and Humate-P® CSL Behring uses cryoprecipitation, Al (OH)3 ad-sorption, glycine precipitation and NaCl precipitation as well as glyci-ne precipitation, dialysis, ultracen-trifugation and sterile fi ltration to achieve a high prion removal capa-city. Recently authorities investiga-ted the risk of transmission of vCJD from plasma-derived products and found it to be extremely minimal.

Prions

* May 2007** For the majority of products.

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Quarantine and inventory hold of source plasma

Quarantine and inventory hold for safety

Figure 3: Quarantine and inventory hold of plasma

Quarantine up to 6 months

New donor

1. Donation(lab test neg.) Inventory hold (min. 60 days)

2. donation(lab test neg.)

+

Plasma for processing

Inventory hold

Regular donor with donation > 6 months since last donation

Regular donor(donation e.g. every 2 weeks)

7. Donation(lab test neg.)

Inventory hold (min. 60 days)

8. Donation(lab test neg.)

9. Donation(lab test neg.)

10. Donation(lab test neg.)

11. Donation(lab test neg.)

12. Donation(lab test pos.)

Inventory hold (min. 60 days)

Inventory hold (min. 60 days)

Inventory hold (min. 60 days)

Inventory hold (min. 60 days)

-

-

-

-

Donor gets informed Donations in inventory are discarded Permanent exclusion of donor

Time

After second donation new donor becomes regular donor

All source plasma donations which have passed the laboratory tests are frozen and stored for a minimum of 60 days. The purpose of this procedure is to be able to react to post donation informa-tion. If a donor is tested positive after the next donation, the pre-

vious donations can be identifi ed and discarded. Such a procedure is very effective when applied to a group of regular donors as is the case with CSL Behring. No plasma pool had to be rejected for the last 6-7 years.

Inventory hold procedures are not regulatory requirements but for CSL Behring it is another part of our commitment to provide the safest possible products (ad-hering to voluntary industry stan-dard „IQPP“ of the PPTA).

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Plasma purifi cation

CSL Behring integrates virus inactivation and elimination procedures in the production process to obtain highly effective and safe products.

Purifi cation

With the process of donor selec-tion and testing it is possible to reduce the potential virus load in the plasma pools to a minimum. To ensure a very high level of safety, virus elimination and in-activation steps are included in the production process. The aim of CSL Behring is to produce pro-ducts with the highest possible degree of safety and quality. Each

coagulation factor has its own characteristics in regards to phy-sio-chemical sensitivity and thus not all products can be treated exactly alike.

CSL Behring integrates purifi ca-tion steps and virus inactivation and elimination procedures to achieve highly effective and safe products.

In the following you will fi nd a short description of CSL Behring’s main manufacturing processes in-volved in the production of plas-ma derived coagulation factors. Please note that not all virus inac-tivation/removal procedures are employed for every product.

Virus elimination stepsa) Plasma fractionation

The recovery of the plasma pro-teins from human plasma is called fractionation. This process is based on the inherent differences of each protein. Fractionation in-volves changing the conditions of the product intermediates (e.g. the temperature, the ionic strength or the acidity) so that proteins become insoluble and

precipitate. The CSL Behring co-agulation factor concentrates are produced using a modifi ed Cohn-fractionation. First the frozen plasma is thawed and pooled. The proteins, which are insoluble under cold conditions (VWF/FVIII and FI (fi brinogen)) are separated by centrifugation and contained in the so called cryoprecipita-

te. In a next step the vitamin K depending coagulation factors (F II, VII, IX and X) are adsorbed from the supernatant and af-ter that factor XIII precipitation takes place through addition of alcohol. Immunoglobulins and al-bumin are precipitated by increa-sing the alcohol concentration.

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Figure 4: CSL Behring plasma fractionation-procedure

Process Intermediates Plasmaprotein Concentrates

Frozen plasma

ThawingCentrifugation Cryoprecipitate FVIII, VWF, Fibrinogen

Cryodepleted plasma

DEAE-Ionexchange-Chromatography

QAE-Ionexchange-Chromatography

PCC, Thrombin, FIX, FX

C1 Inhibitor

Adsorbed cryodepleted plasma

Ethanol-precipitation (8 %)Centrifugation

8 % precipitate (fraction I) FXIII

8 % supernatant

Affi nity-chromatography

Antithrombin III

Adsorbed 8 % supernatant

Ethanol-precipitation (25 %)Centrifugation

25 % precipitate (fraction II + III)

Immunoglobulin

25 % supernatant

Ethanol-precipitation (40 %)Centrifugation

40% precipitate (fraction IV+V) Human Albumin, API

40 % supernatant

Plasma fractionation

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b) Monoclonal purifi cation and affi nity chromatography

Monoclonal purifi cation

1. Addition of factor containing solution

2. Column of monoclonal antibodies with affi nity to a specifi c coagulation factor

3. Forming of coagulation factor/antibody complexes on the column and washout of potential contamination

anti-bodies

Ca++

Ca++

Ca++

Ca++

Ca++

Ca++

Ca++

Ca++

Ca++

4. After washout the specifi c coagulation factor is eluted

5. Purifi ed factor concentrate

Monoclate-P®/Mononine® Virus eliminatione.g. Mononine®

5,8 - ≥ 8,1 log10*

Antibodies against VWF and FIX

buffer

contaminant

Figure 5: Scheme of monoclonal purifi cation

The monoclonal purifi cation of Mononine® and Monoclate-P® is an innovation of CSL Behring that utilises the affi nity of mo-

noclonal antibodies to VWF or FIX respectively. Complexes with the factor proteins are formed and bound to the column and by

rinsing with a buffered solution, viruses and plasmaprotein impu-rities are eliminated.

* log10 = common (base 10) logarithm

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Beriate® P/Berinin® PBeriplex® P/NFibrogammin® P

Virus elimination

Ionexchange chromatography

buffer

In the case of ionexchange chro-matography for the products Beriate® P, Berinin® P, Fibrogam-min® P or Beriplex® P/N different ionexchange chromatography gelsare used to bind the coagula-

tion factors due to their different ionic characteristics. Subsequent rinsing with buffer eliminates plasmaprotein impurities and vi-rus particles.

This purifi cation process effec-tively reduces the viral load. For in-creased safety CSL Behring imple-mented additional inactivation/elimination steps (e.g. pasteurisa-tion, virus fi ltration).

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c) Virus fi ltration (Beriplex®P/N, Mononine®)

Principle of virus fi ltration

Elimination of virusese.g. Beriplex®P/N: HIV ≥ 7,3 log10, BVDV 4,4 log10

75 nm - Filter

35 nm - Filter

Product after virus fi ltration

Basic material with virus particles

Figure 6: Virus fi ltration – example of two fi lters with different pore sizes

The principle of virus fi ltra-tion involves the use of fi l-ters with a defi ned small pore size removing viruses with a diameter larger than the pore

size. The main advantage of this method is that a complete elimi-nation of larger viruses can be achieved without any changes in the quality of the derived pro-

duct. Virus fi ltration may only be used if the desired protein has a smaller size than the pores in the fi lters.

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Virus inactivation

Pasteurisation – an effective technology

Pasteurising proteins

Solution of proteins

Addition of stabilising agents

Pasteurisation (10 h or more depending on the product; 60° C in aqueous solution)

Elimination of non-toxic stabilising agents

Continued production process

Pasteurisation

Pasteurisation(10 h or more depending

on the product; 60° C in aqueous solution)

Factor VIII

Pasteurisation

non- enveloped virus

inactivated virus

enveloped virus

inactivated virusFactor VIII

Figure 7: The effi cacy of pasteurisation

Figure 8: Pasteurisation – fl ow scheme

The pasteurisation of coagulation factor concentrates is mastered to perfection by CSL Behring. It is used for almost all CSL Behring plasma-derived coagulation fac-tors. The relatively simple princip-le is based on heating the products for 10 hours or more depending on the product at a temperature of 60° C in aqueous solution. The addition of stabilising agents such as sucrose and glycine or neutral

salt protects the plasma proteins against denaturation.

The S/D treatment only inactiva-tes non-enveloped viruses. CSL Behring however utilises pasteu-risation. Its main advantage is the inactivation of a wide range of enveloped as well as non-envelo-ped viruses.

More than 8 billion VWF:RCo IUs of Haemate P/Humate-P, the fi rst pasteurised VWF/FVIII concentra-te, have been infused over the past 25 years and not a single case of proven virus transmission has been documented. In addition, Haema-te P/Humate-P pasteurisation has also led to the effective inactiva-tion of the following emerging viruses: infl uenzavirus, coronavi-rus, bunyavirus and fl avivirus.

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No documented case of virus transmission in the 25 year history of Haemate® P/Humate® P, the fi rst effectively virus inactivated VWF/FVIII.

Validation of virus reduction steps

Laboratory experiments are per-formed to evaluate the effi cacy of virus reduction. The different manufacturing steps are run in a laboratory scale. Through the addition of a defi ned amount of virus it is possible to quantify the reduction of virus after individu-

al procedures. The major test-vi-ruses as requested by the autho-rities are: HIV, BVDV (bovine viral diarrhoea virus) as a model for hepatitis C and G virus, herpes virus, HAV, and CPV (canine par-vovirus) as a model for parvovi-rus B19. In all laboratory tests it

is necessary to maintain the con-ditions of relevant parameters in the manufacturing processes to obtain correct results. All tests are performed for each virus se-parately and repeated to provide valid data.

For all plasma derived products CSL Behring validates the manufacturing processes for the capacity of elimination and inactivation of viruses. For every product effective virus reduction (= inactivation and removal) is being performed and all products of CSL Behring have high total reduction factors.

Validation process

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Mean value of total reduction factors (log10)

Enveloped viruses Non-enveloped viruses

HIV BVDV Herpes virus HAV Parvoviruses

Beriate® P ≥ 10.0 ≥ 12.4 8.7 6.6 ≥ 8.7

Berinin® P ≥ 15.6 ≥ 11.9 ≥ 12.6 7.6 4.4

Haemate® P ≥ 12.1 ≥ 12.7 11.6 9.2 ≥ 10.2

Monoclate® P ≥ 13.7 ≥ 7.3 ≥ 9.2

Mononine® ≥ 11.7 ≥ 12.2 ≥ 15.5 ≥ 5.1 ≥ 12.0

Beriplex® P/N ≥ 22.8 ≥ 15.2 ≥ 23.1 6.1 6.3

Fibrogammin® P ≥ 17.2 ≥ 13.3 ≥ 14.6 9.0 ≥ 7.3

Haemocomplettan® P ≥ 12.4 ≥ 12.7 ≥ 9.1 ≥ 7.7 ≥ 8.4

Kybernin® P ≥ 15.1 ≥ 15.7 ≥ 14.8 ≥ 5.4 10.4

Table 4: Total reduction factors for different CSL Behring products. Virus reduction for Parvoviruses includes Parvovirus B19 inactivation, when pasteurisation experiments were performed.

Quality control

CSL Behring conducts quality con-trol measures after every single process step and additionally at the end of the manufacturing pro-cess. This repeated control ensures the consistent high quality of the products.

CSL Behring‘s safety processes were among the fi rst to be certifi ed by the PPTA‘s internationally recogni-sed safety program, QSEAL. Regular inspections by health authorities ensure compliance with global in-dustry standards.

According to the German Paul-Ehrlich Institute the benchmark for total reduction factor for en-veloped viruses is > 10 log10 levels

and for non-enveloped viruses > 6 log10 levels. In almost all cases CSL Behring exceeds these bench-marks. Table 4 shows the CSL Beh-

ring levels of overall reduction factors for enveloped as well as non-enveloped viruses.

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23

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4. Recombinant products

Next to purifi cation from human plasma, coagulation factors can be provided through recombi-

nant technology. CSL Behring has contributed to several processes involved in recombinant manu-

facturing of FVIII such as Helixa-te® NexGen/FS.

The rFVIII manufacturing phases

Phase I: Fermentation • Cell bank • Propagating • Cultivation

Phase II: Purifi cation • Purifi cation by monoclonal AB adsorption* • Virus inactivation by S/D treatment

Phase III: Formulation • Formulation** • Final processing

Manufacturing process

Figure 9: Manufacturing process of recombinant factors

* The immuno chromatography was orignally developed by Scripps Clinic, La Jolla, California for what is now CSL Behring.

** The albumin free sucrose for-mulation was developed by Behringwerke AG.

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Fermentation

Baby Hamster Kidney (BHK) cells are used for the expression of human factor VIII. This cell line is well investigated and has been cultured for many years. After transfecting the factor-VIII gene into the cells, they have the abili-ty to synthesise human factor VIII.

For the industrial production of FVIII these cells are cultivated in a bioreactor containing fresh medium. All cells are genetically identical and therefore produce identical FVIII molecules which are released into the medium. Continuously the factor VIII con-

taining harvest solution is removed and purifi ed to the fi nal rFVIII-product.

Recombinant FVIII-Helixate® NexGen/FS

Plasmid integrated into DNA of baby hamster kidney cells 150 copies/cell

Harvesting of human FVIII

Virus plasmid with total human F VIII gene integrated

Cell culture produces human FVIII and transports it to medium

Terminator Coding sequence

for F

VIII

Promoter

DH

PR

8o KD B 9

0 K

D

pAML 3R8c1

Figure 10: Production steps of recombinant FVIII

Procedure is described on page 16.

Monoclonal purifi cation

26

Inactivation by using S/D

Solvent/detergent (S/D) virus inactivation

Treatment with the combination of a solvent and a detergent

Factor VIII

non- enveloped virusenveloped virus

non- enveloped virus

inactivated virus

Factor VIII

Figure 11: S/D virus inactivation

Due to the presence of retrovirus-like particles in rodent cell lines and in order to reduce adventi-tious viruses potentially present in the fermenter harvest, virus reduction methods are included also in the manufacturing process of recombinant proteins. The S/D

virus inactivation is implemen-ted into the manufacturing pro-cess, as a well known method to effectively inactivate enveloped viruses due to a destruction of the lipid envelope. Without the envelope, viruses cannot infect target-cells and further virus rep-

lication is inhibited. The integrity of the therapeutic proteins is not affected.

Validation studies show that S/D combined with the manufac-turing‘s virus reduction capacity is suffi cient.

Albuminfree formulation

Formulation of recombinant FVIII

• Formulated with sucrose • Reduction of foreign proteins

Reduced risk of anaphylaxis

Figure 12: Formulation process

This formulation process was de-veloped by CSL Behring. Helixate® NexGen/FS is stabilised with su-crose. It is one more step to mi-

nimise the use of foreign prote-ins. Until now 2,5 billion IUs of Helixate® NexGen/FS have been infused and there has never been

a case of transmission of an infec-tious agent reported.

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5. Summary

The elimination and inactivation of viruses in the manufacturing process of products is achieved by a series of measures. It starts with the selection of donors and tes-ting of donations, proceeds with purifi cation, virus inactivation and elimination and is completed by multiple measures of quality control and quality assurance.

The pasteurisation used by CSL Behring for most of the products is a very effective elimination procedure for enveloped and non-enveloped viruses and has been in use for > 25 years. Pro-ducts may have a further virus in-activation/elimination step if the quality of the individual product is not jeopardised. For products incompatible with pasteurisation

combinations of other virus inac-tivation/elimination steps are im-plemented.

6. Abbreviations

BVDV Bovine Viral Diarrhoea Virus

CJD Creutzfeldt-Jakob-Disease

DNA Deoxyribonucleic acid

F Factor

HAV Hepatitis A Virus

HBV Hepatitis B Virus

HCV Hepatitis C Virus

HIV Human Immundefi ciency Virus

HSV Herpes Simplex Virus

NAT-PCR Nucleic Acid Amplifi cation Technology-Polymerase Chain Reaction

PPTA Plasma Protein Therapeutics Association

PRV Pseudorabies Virus

RNA Ribonucleic acid

S/D Solvent-detergent treatment (virus inactivation method)

TF Tissue Factor

VWD von Willebrand Disease

VWF von Willebrand Factor

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7. Literature

1. Abshire TC et al.: Sucrose formulated recombinant human antihemophilic factor VIII is safe and effi cacious for treatment of hemophilia A in home therapy. Thromb Haemostas 2000; 83(6): 811-816

2. Blood Diagnostics: Virus safety info-mations: http://www.ivig.com/Viral_safety.html

3. Committee for Proprietary Medicinal Products (CPMP):

Validation of virus removal and in-activation procedures. Biologicals 19 (1991): 247-251

Guidelines for Minimizing the risk of transmitting agents causing spongi-form encephalopathy via medicinal products. Biologicals 20 (1992): 159-164

Guidelines for medicinal products de-rived from human blood and plasma. Biologicals 20 (1992): 159-164

4. CSL Behring Data on fi le

5. Gröner A: Purity, Activity and virus safety of a pasteurized antithrombin concentrate. Semin Thromb Hemostas 2002; 28 (Suppl. 1): 79-85

6. Heimburger N. From cryoprecipitate to virus safe high purity concentra-tes. Semin Thromb Hemostas 2002; 28 (Suppl. 1): 25-31

7. Hoffman M and Monroe DM. A cell-based model of hemostasis. Thromb Haemost 2001; 85: 958-65

8. Johannsen R et al.: Integrated safety system of plasma derived medication. Die gelben Hefte Jg. 37 1997: 161-171

9. Kreuz W et al.: Virus safety of pasteu-rized clotting factor concentrates. Se-min Thromb Hemostas 2002; 28 (Sup-pl. 1): 57-61

10. Mannucci PM. Coagulation factors: The State of Safety; XVII ISTH Congress Satellite Symposion Washington 1999. Medizinische Verlagsgesellschaft Um-welt & Medizin mbH

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