validation virus removal 092600 kw

Validation of Virus RemovalValidation of Virus Removal

Keith O.Webber, Ph.D.Keith O.Webber, Ph.D.Deputy Director, Div. of Monoclonal Deputy Director, Div. of Monoclonal

AntibodiesAntibodiesOTRR/CBER/FDAOTRR/CBER/FDA

PDA/FDA MeetingPDA/FDA MeetingSeptember 26, 2000September 26, 2000

SCOPE - ProductsSCOPE - Products

Specified BiologicsSpecified Biologics Monoclonal AntibodiesMonoclonal Antibodies rDNA-derived productsrDNA-derived products

Traditional BiologicsTraditional Biologics Some Blood-productsSome Blood-products Some VaccinesSome Vaccines

SCOPE - MethodsSCOPE - Methods

Column ChromatographyColumn Chromatography

Nano-Filtration (Viral Filtration)Nano-Filtration (Viral Filtration)

Other MethodsPrecipitationCentrifugation

ExampleExample

Production FermentorProduction Fermentor

Protein A affinity chromatographyProtein A affinity chromatography

Anion Exchange ChromatographyAnion Exchange Chromatography

Virus filtrationVirus filtration

Gel FiltrationGel Filtration

Formulation and FillFormulation and Fill

GENERALITIESGENERALITIES

Validation conditions Validation conditions mustmust be representative of be representative of the actual manufacturing process. the actual manufacturing process.

Validation should use virus-spiking studies. Validation should use virus-spiking studies.

Viral clearance studies should Viral clearance studies should notnot be done in be done in the production facility.the production facility.

Validations should be done in duplicate.Validations should be done in duplicate.

SCALE-DOWNSCALE-DOWN

Scale Down:Scale Down:» Allows validation to be performed in testing labsAllows validation to be performed in testing labs

» Maintains high titers of the spiking virusMaintains high titers of the spiking virus

» Must be done appropriatelyMust be done appropriately

CHROMATOGRAPHYCHROMATOGRAPHY

Parameters that should be representative of Parameters that should be representative of commercial-scale manufacturing:commercial-scale manufacturing:

– CHROMATOGRAPHY MEDIUMCHROMATOGRAPHY MEDIUM– COLUMN BED HEIGHT COLUMN BED HEIGHT – LINEAR FLOW RATELINEAR FLOW RATE– BUFFER COMPONENTS AND CONCENTRATIONSBUFFER COMPONENTS AND CONCENTRATIONS– pHpH– TEMPERATURETEMPERATURE– PRODUCT LOADPRODUCT LOAD

CHROMATOGRAPHYCHROMATOGRAPHY

and…and… the chromatographic profile and product recovery of the chromatographic profile and product recovery of

the scale-down process should be comparable to the scale-down process should be comparable to that of the manufacturing process.that of the manufacturing process.

FILTRATIONFILTRATION

Parameters that should be representative of Parameters that should be representative of commercial-scale manufacturing:commercial-scale manufacturing:

– VISCOSITYVISCOSITY– VOLUME PER cmVOLUME PER cm22 OF FILTER AREA OF FILTER AREA– IONIC STRENGTHIONIC STRENGTH– TEMPERATURETEMPERATURE– pHpH– PROTEIN COMPOSITION & CONCENTRATIONPROTEIN COMPOSITION & CONCENTRATION

FILTRATIONFILTRATION

and…and…the product recovery of the scale-down the product recovery of the scale-down process should be comparable to that of the process should be comparable to that of the manufacturing process.manufacturing process.

CHOOSING VIRUSESCHOOSING VIRUSES

The aims of viral validation studies are:The aims of viral validation studies are:» to provide evidence that the production process to provide evidence that the production process

will effectively remove viruses which are either will effectively remove viruses which are either known to contaminate the starting materials, or known to contaminate the starting materials, or which could conceivably do so, andwhich could conceivably do so, and

» to provide indirect evidence that the production to provide indirect evidence that the production process might remove novel or unpredictable process might remove novel or unpredictable virus contamination.virus contamination.


The clearance validation should include:The clearance validation should include:» relevant virusesrelevant viruses that may be anticipated to occur that may be anticipated to occur

in the system in the system

» specific model virusesspecific model viruses that are physically and that are physically and chemically similar to relevant viruseschemically similar to relevant viruses

» non-specific model virusesnon-specific model viruses that represent the that represent the extremes of virus propertiesextremes of virus properties


Examples of model viruses:Examples of model viruses:Retrovirus:Retrovirus: X-MuLV X-MuLV

Small non-enveloped virus:Small non-enveloped virus: SV40, parvovirus, polio virus 1 SV40, parvovirus, polio virus 1

Medium to large enveloped RNA virus:Medium to large enveloped RNA virus: Parainfluenza, Parainfluenza, Sindbis virusSindbis virus

Medium to large DNA virus:Medium to large DNA virus: HSV-1, pseudorabies virus HSV-1, pseudorabies virus

PRECAUTIONSPRECAUTIONS

Avoid aggregation of spiking virusAvoid aggregation of spiking virus Use small volumes of spiking virus in order to retain the Use small volumes of spiking virus in order to retain the

sample composition.sample composition. Assess the potential for assay interference by the product or Assess the potential for assay interference by the product or

buffers. buffers. Use control assays in parallel to assess the loss of infectivity Use control assays in parallel to assess the loss of infectivity

due to dilution, concentration, filtration, or storage.due to dilution, concentration, filtration, or storage. Include controls to demonstrate the effect of procedures Include controls to demonstrate the effect of procedures

used solely to prepare the sample for assay.used solely to prepare the sample for assay.

General ProcedureGeneral Procedure

Add high Titer SpikeAdd high Titer Spiketo Sample Loadto Sample Load

•Titer Sample Load Titer Sample Load •Take Hold Control sampleTake Hold Control sample

•Titer Hold ControlTiter Hold Control•Titer Column Flowthrough Titer Column Flowthrough •Titer Eluate SampleTiter Eluate Sample

Virus Assay Methodology

Infectivity is the standard method for Infectivity is the standard method for clearance studies.clearance studies.

PCR assays and Polymerase Enhanced PCR assays and Polymerase Enhanced Reverse Transcriptase assays are being Reverse Transcriptase assays are being developed.developed.

Evaluation of ResultsEvaluation of Results

Clearance factors from sequential Clearance factors from sequential orthogonal processes may be combined to orthogonal processes may be combined to give a Cumulative Clearance Factorgive a Cumulative Clearance Factor

Orthogonal processes are those that clear Orthogonal processes are those that clear virus by independent modes of action (e.g., virus by independent modes of action (e.g., Anion Exchange chromatography and Anion Exchange chromatography and Cation-Exchange chromatography)Cation-Exchange chromatography)


Beware of including bothBeware of including both

a low-pH treatment step and a low-pH treatment step and

a chromatography step that uses elution at a chromatography step that uses elution at a low pHa low pH

in the Cumulative Clearance Factor.in the Cumulative Clearance Factor.


The Cumulative Clearance Factor must be The Cumulative Clearance Factor must be substantially greater than the estimated substantially greater than the estimated number of virus particles in a volume of the number of virus particles in a volume of the starting material required to produce a starting material required to produce a human dose of the drug.human dose of the drug.


The number of virions per mL of starting The number of virions per mL of starting material should generally be estimated by material should generally be estimated by Transmission Electron Microscopy.Transmission Electron Microscopy.

One “dose” of product is generally One “dose” of product is generally considered to be the total regimen of drug.considered to be the total regimen of drug.

e.g., 1 mg/week x 4 weeks = 4 mg dosee.g., 1 mg/week x 4 weeks = 4 mg dose

ExampleExample

Product Product

Protein A affinity chromatographyProtein A affinity chromatography

Anion Exchange ChromatographyAnion Exchange Chromatography

Virus filtrationVirus filtration

Gel FiltrationGel Filtration

Formulation and FillFormulation and Fill

Homologous ProductHomologous Product

Generic and Modular ValidationGeneric and Modular Validation

A generic validation study demonstrates A generic validation study demonstrates virus removal or inactivation by a process virus removal or inactivation by a process using a using a model productmodel product and allows the use and allows the use of that process with an homologous of that process with an homologous product without the need to revalidate.product without the need to revalidate.


Generic viral clearance validation is Generic viral clearance validation is applicable to situations when the applicable to situations when the purification process of a product is the purification process of a product is the same as a process that has already been same as a process that has already been validated for an homologous product.validated for an homologous product.

ChromatographyChromatography

The two processes must have the same:The two processes must have the same: -chromatography medium,-chromatography medium,-column geometry,-column geometry,-equilibration buffers,-equilibration buffers,-load composition & concentration,-load composition & concentration,-elution buffers,-elution buffers,-elution parameters,-elution parameters,-wash procedure.-wash procedure.

Virus FiltrationVirus Filtration

The two processes must have the same:The two processes must have the same: type of virus filtertype of virus filter solution characteristicssolution characteristics

VOLUME PER cmVOLUME PER cm22 OF FILTER AREA OF FILTER AREA IONIC STRENGTHIONIC STRENGTH TEMPERATURETEMPERATURE pHpH PROTEIN COMPOSITION & CONCENTRATIONPROTEIN COMPOSITION & CONCENTRATION


Example of Homologous Products:Example of Homologous Products:Monoclonal Antibodies of the same Monoclonal Antibodies of the same species, class, and subclass and species, class, and subclass and derived from the same source (e.g., derived from the same source (e.g., ascites, tissue culture, etc.) and cell ascites, tissue culture, etc.) and cell substrate.substrate.


NOTE:NOTE:

GENERIC VALIDATIONS ARE GENERIC VALIDATIONS ARE NOTNOT APPLICABLE TO HUMAN-DERIVED APPLICABLE TO HUMAN-DERIVED PRODUCTS OR PRODUCTS THAT MAY BE PRODUCTS OR PRODUCTS THAT MAY BE CONTAMINATED WITH HUMAN CONTAMINATED WITH HUMAN PATHOGENS.PATHOGENS.

Post-Approval ChangesPost-Approval Changes

Whenever a change is made in the Whenever a change is made in the production or purification process, the production or purification process, the effect of that change on the viral clearance effect of that change on the viral clearance should be considered and the system re-should be considered and the system re-validated as needed.validated as needed.

Additional ReadingAdditional Reading ICH Guidance on Viral Safety Evaluation of Biotechnology Products Derived from Cell Lines of

Human or Animal Origin,1998 (www.fda.gov/cber/guidance/)

Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use, 1997 (www.fda.gov/cber/guidance/)

Guideline on General Principles of Process Validation, 1987 (www.fda.gov/cder/guidance/pv.htm)

Reviewer Guidance: Validation of Chromatographic Methods, 1994 (www.fda.gov/cder/guidance/)

Validation of the Purification Process for Viral Clearance Evaluation, 1997, A.J. Darling, in Biopharmaceutical Process Validation, G. Sofer and D.W. Zabriskie, editors. Marcel Dekker, Inc (New York)

http://www.fda.gov/cder/guidance/

http://www.fda.gov/cder/guidance/

GuidanceGuidance

CBER Office of Therapeutics:CBER Office of Therapeutics: 301-827-5101301-827-5101

CBER Office of Vaccines:CBER Office of Vaccines: 301-827-3070301-827-3070 CBER Office of Blood:CBER Office of Blood: 301-827-3524301-827-3524

Obtaining DocumentsObtaining Documents

Fax Fax (888) CBER-FAX(888) CBER-FAX

Internet http://www.fda.gov/cber/guidelines.htmInternet http://www.fda.gov/cber/guidelines.htm

E-mail [email protected] [email protected]

Mail Mail Training and Manufacturer Assistance Training and Manufacturer Assistance (HFM-40) CBER/FDA(HFM-40) CBER/FDA1401 Rockville Pike1401 Rockville Pike Rockville, MD 20852-1448Rockville, MD 20852-1448

validation virus removal 092600 kw

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