Viral Inactivation of Blood Products

Soheir Adam, MD, MSc., MRCPathAsst. Professor & Consultant H t l i tHematologist

Human and animal Blood derived medicinal products & related in vitro diagnostic devices

Animal- derived sera Anti-rabiesA ti ( k / i )

Human blood derived productsBlood components (red cells, Anti-venoms (snake/scorpion)

Anti-tetanus toxinsAnti-diphteria toxins

p ( ,platelets, plasma) Blood Coagulation FactorsImmunoglobulins

A i h i i B Anti-botulism toxinsAnti-hepatitis BAnti-rabiesAnti-tetanusAnti-rhesus (anti-D)

Fibrin sealantAlbumin

Other related products

Anticoagulant & fibrinolysis Albumin

In vitro diagnostic devices for the control of q alit and safet of blood

Anticoagulant & fibrinolysis biological therapeutic products

In vitro diagnostic devices for the control of quality and safety of blood derived medicinal products including in vitro diagnostic devices

Th i CThe main Concern:

encouragement of

voluntary & unpaid bloodvoluntary & unpaid blood donationsdonations

Quality and Safety: Q y yBiological Products

Blood collection & Blood collection & Plasma quality & safetyPlasma quality & safetyStarting materialStarting material

FractionationFractionation

q y yq y y

Fractionation Fractionation Technology/ Viral Technology/ Viral inactivation /Removal inactivation /Removal

Production Production ProcessProcess

ProceduresProceduresProcessProcess

Product characteristics:Product characteristics:Bulk &Bulk & Formulated Formulated ProductProduct

Final product Final product consistencyconsistency

Product Product

Transfusion TransmissibleTransfusion Transmissible Diseases

Hepatitis BHepatitis CHepatitis CHIV 1&2HTLV 1&11HTLV 1&11SyphilisCMVCMVBacterial contaminationvCJDWest Nile VirusParasites

Residual risks of major TTIs*

1993-1996 2000-2002WPWP Serology WP NATdays days

HIV 22 1: 3 000 000 11 1: 5 000 000HIV 22 1: 3 000 000 11 1: 5 000 000HCV 70 1: 500 000 10 1: 5 000 000HBV 68 1: 135 000 45 1: 250 000**

* Schreiber et al NEJM 1996 ;334:1685-90 ** serology

Transfusion Safety

Careful selection of donorsTestingViral inactivation /recombinant productsAppropriate blood useAppropriate blood useHaemovigilance

Why Viral Inactivation

The growing list of blood- Borne th i t i bilit t d ipathogens is outpacing ability to devise

and implement new screening tests

Plasma

Platelets & leucocytesPlatelets & leucocytes

Red Cells

PlateletsPlatelets

Platelet Production

2nd ti i2nd separation in an extractor

Leucoreduction by filtration

D S iDonor Screening

Donor TestingDonor Testing

Viral removal/ inactivation

Quarantine

P d t lProduct release

Prinicples of Collection and Manufacturing Process

Pl T tiPlasma Testing

Plasma tested for HBsAg

Plasma pools tested forHBsAg

Anti- HCVAnti HIV

tested forHBsAgAnti- HCVAnti-HIV

ALTPCR for HCV

Anti- HCVAnti-HIVPCR for HCVPCR for HCV,

HBV, HIV, Parvovirus B19

PCR for HCV, HBV, HIV, Parvovirus B19

Nucleic Amplification TestingNucleic Amplification Testing(NAT)

HCV NAT SNBTS Nov 99HIV NAT Sept 01Approx 950 000 screened ppNo HCV RNA, anti-HCV neg detectedNo HIV RNA anti HIV neg detectedNo HIV RNA, anti-HIV neg detected

UK 3: 3,500,000 HCV RNA pos

Product Safety

Full Traceability From Donor To Final ProductNotification System For Advising Of Post DonationNotification System For Advising Of Post Donation InfectionsValidated Virus Elimination Step(s)Validated Virus Elimination Step(s)Control Of Process To Prevent Re-contamination After Virus Elimination SteppClinical Trial DataPost Marketing Surveillanceg

Vi l S f tViral SafetyResearch into effective methods for inactivation or removal of viruses from blood products

Chemical Inactivation Inactivation by heatRemoval by virus filter (nanofiltration)Removal by alcohol precipitationRemoval by alcohol precipitation

Vir s Inacti ation and Remo alVirus Inactivation and RemovalD i th d th t ill l ti l i ti t d/• Devise methods that will selectively inactivate and/or remove viruses without undue product damage/loss.

St d l t t t i i l d d• Study relevant test viruses in scaled-down process.

• Ensure that method is capable of giving the degree of virus inacti ation/remo al req iredinactivation/removal required.

Common Virus ClearanceCommon Virus Clearance Methods

Virus inactivation: Virus removal

Chemical:organic solvents; pH extremes;

Precipitation:ammonium sulfate etc; p ;

solvent detergent; alcoholPhysical: Heat treatment (dry heat or

ammonium sulfate etc.Chromatography: ion exchange; gel filtration; (dry heat or

pasteurization) affinity; reverse phaseMembrane filtration:Omega, Planova, DV50Omega, Planova, DV50

Criteria for An Effective VirusCriteria for An Effective Virus Clearance Step

Si ifi t i l killSignificant viral killReproducible and controllable at process

l d d l bl t th l b tscale and model-able at the laboratory scaleSh ld h i i l i t d tShould have minimal impact on product yield and activityN t t ti l t iNot generate neo-antigens or leave toxic residues

Vi S f tVirus Safety

Cold ethanol fractionationH4/ i i i ti tipH4/pepsin virus inactivation

effective against enveloped viruses e.g. HIV, Hep B and Cand non-enveloped viruses e.g. Hep A

Red Blood Cells

Hb has phostoabsorptive effects.Light – independent compounds have been devised.S- 303, PEN110 have no adverse effects.

Fresh Frozen Plasma

PsoralenRiboflavinSolvent – detergent plasmag p

Large pools : 250,000 donors/ batchThrombotic complicationsThrombotic complications

Platelets

No approved method of viral i ti ti f l t l tinactivationof plateletsS- 59 is effective but less effective clinically than untreated platelets.

Pooled screened human p lasma

C ld th l f ti ti t i ldCold ethanol fractionation to yieldfraction II

Remove process ethanolRemove process ethanol

pH4/pepsin VI step

Sterilisa tion, aseptic dispensing

Product returned to a neutral pH

and freeze drying

SNBTS Intravenous Immunoglobulin

ConclusionConclusion

Manufacturing processes for blood g pderived products should contain two effective steps for premoval/inactivation of viruses

At least one step should be effective against non-enveloped virusesagainst non-enveloped viruses

ConclusionConclusion(cont.)

At least one stage in a production processAt least one stage in a production process must inactivate rather than remove viruses

A single step having a large effect gives more f i l f t th l tassurance of viral safety than several steps

having the same overall effect