validation in biotech facilities: what ? why ?...

Validation in Biotech Facilities: What ? Why ? How?

Dr. PK Yegneswaran

2 2

Presentation Outline

� Validation Overview

� Cleaning Validation

� Process Validation

� Sterilization Validation

� Citation Examples

� Regulatory References

3 3

Typical Project Schedule

1 2 3 4 5 6

Scope

Design

Procurement

Construction

IQ/OQ

Startup / Validation

100%

YEAR

APPROVAL

% S

pe

nt

Phase III

APPROVAL

4 4

Typical Post OQ Schedule

Cleaning Dev.

Sterilization Dev.

Practice Lots

IQ/OQ/Facility/Utility Qualification

Validation Lots

YEAR

APPROVAL

Phase III

APPROVAL

1H Y4 2H Y4 1H Y5 2H Y5

Cleaning Valdn.

Sterilization Valdn.

File Licens

e

5

Validation Overview

[To establish] documented evidence which provides a

high degree of assurance that a specific process

will consistently produce a product meeting pre-determined specifications and quality attributes.

(FDA, May 1987)

6 6

Validation Overview

� Why validate ?

– The FDA requires that we validate all of our

systems and processes according to 21 CFR part 211

– Improves our understanding of our manufacturing

processes

– Right thing to do !

7 7

Why Validate?

� Consistent yield &

quality

� Rapid decisions when

mishaps occur

� Fewer discards

� Less time hosting government agencies,

more time

manufacturing

= to ensure that the

output is consistent;

first time, every time!!

8 8

What do we Validate ?

� Process

� Cleaning

� Sterilization

� Filters

� Containers

� Assays

9 9

How do we Validate ? Cleaning, Process, Sterilization etc.

� Define Critical Process Parameters, Critical Quality Attributes

� Develop protocol describing validation studies

– Consider fractional study approach for cleaning, sterilization

� Execute studies

� Address deviations

� Compile report

� Review / Approve report

� GMP Documentation all the way=.

This process applies to all validation

10 10

Definitions

Critical Process Parameter (CPP):

An input variable that must be controlled within a specified range to ensure success.

Critical Quality Attribute (CQA):

An output parameter from a unit operation that must be within a specified range to demonstrate control, consistency, and acceptable product quality.

CPP CQA

Ionic Strength Ion Exchange Yield Column Load Chromatography Purity

Flow rate Cleaning Conductivity

Temperature TOC

Concn.

Sat. Steam Sterilization BIs

Time

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Style-ogen® Facility Built

IQ/OQ Complete What Next ?

• Sterilization Validation • Development, validation studies

• Practice Lots • Define validation parameters for process, cleaning, cleaning

validation

• Validation Lots • Process validation, cleaning validation

• File license

• Pre-Approval Inspection

• Facility / Product approval

12 12

Typical Post OQ Schedule

Cleaning Dev.

Sterilization Dev.

Practice Lots


Validation Lots

YEAR

APPROVAL

Phase III

APPROVAL

1H Y4 2H Y4 1H Y5 2H Y5

Cleaning Valdn.


File Licens

e

13 13








14 14

Cleaning Validation

� Cleaning Validation overview

� Cycle development for Style-ogen®

equipment

� Validation of cleaning cycles

15 15

Equipment cleaning validation is the process

of establishing documented evidence that a

particular cleaning procedure will consistently

reduce equipment surface residuals to a predetermined acceptable level.

“Residuals” are any product, degradate,

intermediate, excipient, raw material/reactant

or cleaning agent that may reside on any equipment surface following processing

and/or cleaning.

What is Cleaning Validation ?

16 16

21 CFR Part 211 Subpart D - Equipment

211.67 Equipment Cleaning and Maintenance

– (a) “Equipment and utensils shall be cleaned,

maintained, and sanitized at appropriate intervals to prevent malfunctions or contamination that would alter

the safety, identity, strength, quality, or purity of the

drug productA.”

17 17

21 CFR Part 211 Subpart F - Production and Process Controls

211.113 Control of Microbiological

Contamination

– (a) “Appropriate written procedures, designed to prevent

objectionable microorganisms in drug products not

required to be sterile, shall be established and

followed.”

– (b) “Appropriate written procedures, designed to prevent

microbiological contamination of drug products

purporting to be sterile, shall be established and

followed.”

18 18

One-Third Of Recent Drug GMP Warning Letters

Cite Cleaning Practices (2002 survey)

Year # of Warning Letters

# of Warning Letters related

to Cleaning

% Related to Cleaning

1999 65 23 35%

2000 71 20 41%

2001 71 20 28%

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Cleaning Validation Where Do I Start?

Validation Strategy

Manufacturing

Process

Cleaning

Process

Mfg. Equipment

& Design

Manufacturing

Process

Cleaning

Process

Mfg. Equipment

& Design

20 20

� Obtain a Process Flow Diagram

� Is Product Inactivation Required ? (Important to ensure inactivation procedure is defined before starting validation)

� Define Applicable Hold Times � Dirty: End of Process to Start of Cleaning � Clean: End of Cleaning to Next Process Use � Sterile: End of SS/SIP to Next Process Use

� What “Residuals” Need to Be Cleaned by the CIP ? � Product (includes degradates, excipients, raw materials, etc.) � Cleaning Agents

� Are the “Residuals” Representative of the Process ? (Important to consider when validating during Practice Runs or Demonstration)

� Is the Equipt. Sanitized or Sterilized after CIP ?

Considerations:

Manufacturing Process

21 21

� Define CIP Type (Manual, Automated)

� Are the Individual Steps of the CIP Procedure Defined ?

� Critical Process Parameters Defined ? (e.g. Flow, Temp.)

� Is the Cleaning SOP available ?

� Does CIP Procedure Clean All Product Contact Surfaces?

(Highlight and Compare Mfg Process to CIP Process on Same P&ID)

� What CIP Cycle Development Work is Planned?

Considerations:

Cleaning Process

22 22

� Are All Equipt/Systems IQ/OQ’d ?

� Define Surface Materials of Construction (Prod. Contact)

� Ensure General CIP Design Principles Followed

� Minimal to No System Deadlegs

� Turbulent Flow Maintained During CIP

� Full Coverage to Vessel During the CIP

� Lines Flooded Completely During the CIP

� Complete System Drainability

� Assess Validation Sample Locations

� Accessibility

� Availability

Considerations:

Mfg. Equipment Design

23 23

� Challenge Strategy: � Hold Times

� Fractional Cycle Approach

� No. of Lots Tested

� Product/Equipment Matrix Required

� Test Methods and Sampling Plan:

� Rinse Sampling

� Swab Sampling

� Visual Inspection

� Analytical Methods

� Assay Selection (Chemical/Micro)

� Assay Validated (Includes Swab Recovery)

� Acceptance Criteria

Defined in a Protocol and Includes:

Validation Strategy

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Overall surface evaluation: final rinse sampling & analysis

USP chemical- purified water methodology

pH 5.0 - 7.0

Conductivity < 3 µS/cm

Endotoxin < 0.25 EU/mL to < 10.00 EU/mL

TOC < 1.0 ppm+(based on system capability)

Bioburden* < 100 cfu/10mL

Product specific varies, typically non-detect

+ Over Negative Control

* Bioburden sampling is performed in systems that are not steamed or sterilized for

bioburden control

Typical Acceptance Criteria

25 25

Back to Style-ogen® :Bulk Portable Tanks CIP

Next Use End of Use Cleaning

24 Hour

Dirty Hold Time 7 Day

Clean Hold Time

� Manufacturing Process:

� Product is Inactivated with Hypochlorite Prior to CIP (SOP)

� Only 100L Portable Tanks Cleaned at Bulk PTS

� One Tank Can Be Cleaned At A Time At Bulk PTS

� Each Tank Can Contain One of the Following Product Soils:

� Active Ingredient “Manny”

� Active Ingredient “Moe”

� Active Ingredient “Jack”

� 25% Sucrose

� Tank is Not SS/SIP After Cleaning

26 26

Style-ogen® - BULK PTS CIP

� Cleaning Process:

� Automated CIP Cycle

� Cycle Steps Defined

� Cleaning SOP Available

� Cycle Development

Planned Concurrent to OQ & Engineering Lots

� Mfg. Equipment & Design:

� Bulk PTS & 100L Style-ogen® Tanks will be IQ/OQ’d

� All Product Contact Surfaces Constructed of Stainless Steel

� Good CIP Design Principles Followed

� Validation Sample Locations Readily Accessible &

Available (Rinse Sample Port)

Hold Time or Cycle

Step Description

Production

Cycle

Dirty Hold Time < 24 hrs.

HWFI Rinse 2 min.

2% Caustic Wash 15 min.

HWFI Rinse 2 min.

1% Acid Wash 5 min.

Final Rinse #1 2 min.





Clean Hold Time < 7 days

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Styleogen® - BULK PTS CIP

� Validation Strategy: Residual

Name

Time to Clean to

Acceptable Level

"Manny" 150

"Moe" 300

"Jack" 150

25% Sucrose 100

Lab Scale Cleanability Results

� How Many Validation Lots?

Hold Time or Cycle

Step Description

Production

Cycle

Validation

Cycle

Dirty Hold Time < 24 hrs. > 24 hrs.

HWFI Rinse 2 min. 2 min.

2% Caustic Wash 15 min. 10 min.

HWFI Rinse 2 min. 2 min.

1% Acid Wash 5 min. 5 min.

Final Rinse #1 2 min. 2 min.




Final Rinse #5 2 min. N/A

Clean Hold Time < 7 days > 7 days

Cleaning Cycle Description � Use a Fractional Cycle Approach

� Caustic Wash Time Reduced 33%

� Final Rinse #5 Eliminated

� Dirty & Clean Hold Times

Challenged During Validation

� 3 Lots “Moe” (Hardest to Clean)

� 1 Lot Equivalency Each Others

28 28

Style-ogen® - Bulk PTS CIP

� Execute Protocol – Document deviations

� Collect samples

� Analyze samples

� Check vs Acceptance Criteria – Pass / Fail / Investigation

� Write Report – Address deviations

� Review / Approve Report – Stakeholders � Include summary in license document

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30

Process Validation - Definition

[To establish] documented evidence which provides a

high degree of assurance that a specific process

will consistently produce a product meeting pre-determined specifications and quality attributes.

(FDA, May 1987)

2001-ms-3767

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� Demonstrate process control and consistency

� Comply with regulatory requirements for

licensure

� Provide assurance that release tests will be met;

the need for some release testing may be

eliminated.

Why Validate the Process ?

32 32

Key Process Variables

Optimization/Process Understanding

Robustness Worst case challenges?

Process Validation at Full-scale

Process

Characterization

Process

Validation

Phase I/II Clinical

process

Lab-scale

process

Manufacturing

process

Lab Scale Validation

Process Validation requires a rational approach

33 33

Characterization vs. Validation

� Characterization

– “Validation” studies at bench-scale using scaled-down

models, if possible.

– Well-documented in Lab notebooks and key technical

reports (no protocol)

– Learning, not “Validating”

� Validation

– Usually at Full-scale in actual process equipment (except

for viral clearance and resin/filter re-use)

– Conducted by Manufacturing under Protocol

– Testing what we already know, NOT EXPERIMENTING!

34 34

Understand Your Process

� Ruggedness

– Multiple lots of raw materials

– Multiple lots of resins/filters

– Explore failure limits at laboratory/pilot scale

� Scaled-down process should reflect full

scale manufacturing performance as

closely as possible so that data generated

are relevant.

35 35

Definitions

Critical Process Parameter (CPP):

An input variable that must be controlled within a specified

range to ensure success.

Critical Quality Attribute (CQA):

An output parameter from a unit operation that must be

within a specified range to demonstrate control,

consistency, and acceptable product quality.

CPP CQA

Ionic Strength Ion Exchange Yield

Column Load Chromatography Purity

36 36

Example-Homogenization Step

Homogenization •Pressure

•Conc. •# of passes

•Temperature •Residence time

•Back-pressure

•Cell breakage

CPPs? CQA

Function in the manufacturing process:

Cell breakage - cell breakage must be ≥≥≥≥ 70% by Hematocrit assay.

• Process knowledge • Scientific rationale

•Tools are simply to provide a basis for discussion

and to facilitate the PV process.

37 37

1. Select CPPs, CQAs

2. Process Validation Protocol

3. Execute

4. Assay

5. Report

6. File

Back to Style-ogen® : Process Validation

38

Process Mapping: Step & Purpose

Fermentation

Thaw/Cell Breakage

Microfiltration/Chromatography 1

UF/Chromatography 2

Sterile Filtration

Dilution/Adjuvant Addition

Antigen Release

Antigen capture

Polishing purification

Sterilization

Dose/ adjuvant

39 39

Detailed Step Description

Chromatography

Step Goal: Primary purification

Equipment constraints: •flow rate •pressure drop

•Cycle time •Column size •Flow distribution

Sampling plan: •Feed •Flow-through

•Product

Characterization •Size •Potency

•Lipid •Carbohydrates •Yield •Purity

Monitoring •Flow-pressure •UV

•Conductivity •HETP

Other parameters: Feed properties/composition, salt concentration, temperature, lot-to-lot feed/resin variability, feed concentration, load

Support Documents •Technical memos:

•Effect of load

•Cleaning/reuse •SOP’s •Batch summaries •Equipment FRS

40 40

� Impact on product quality

– does the parameter have an impact on a CQA?

� Controllability

– how easy is the parameter to control?

� Recovery potential

– is there a redundant downstream step?

Use tools such as Criticality Index Analysis

Select CPPs, CQAs – Factors to Consider

41 41

Example of a Criticality Index Analysis

Cell breakage

Enzyme

treatment

Microfiltration

42 42

Process Validation Protocol

� CPPs, CQAs w/ acceptance criteria – Background / rationale for ranges

� How will they be sampled / monitored ?

� How many validation lots ?

� How will deviations be handled ?

Define Roles and Responsibilities

Manufacturing, Quality, Technology

43 43

Process Validation Protocol

Step Goal CPPs CPP

Range

How

controll

ed

CQA Samples CQA

Range

Methods

Ferment

ation

High

cell

density

pH

Temp

7.0± 0.5 DCS Final

Glucose

Concn.

Broth –

final time

point

1 – 3

g/L

Analytical

methd

SOP XYZ

44 44

Style-ogen® - Process Validation

� Complete 3 Validation Lots

� Obtain, Analyze data

� Address deviations �Transient deviations

�Equipment malfunctions

� Additional lots if needed

� Complete / approve report

� Include in license

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46 46

Sterilization Validation

� Sterilization Validation overview

� Validation of sterilization cycles (Protocol,

Acceptance criteria,=.)

47 47

Steam Sterilization

� Cell death by protein denaturation

� Simple, reliable & economical

� Spores are more resistant than cells

� Spores ~100x more resistant to dry heat

than steam

� Typical cycle: >121ºC for 5-45 minutes

� Saturated steam is critical!

48 48

Kinetics of Microbial Death

� Generally observed to be first-order

kinetics

� Non-logarithmic behavior is known

� Kinetic models

)( kt

o

oo

eNN

ttatNN

kNdt

dN

−=

==

=−

49 49

Kinetics of Cell Death

50 50


� Logarithmic decline most applicable to

vegetative cells

� Spores can show non-log rates

– Spore germination

– Sequential events for death

SSRR

S

RR

R

D

k

S

k

R

NkNkdt

dN

Nkdt

dN

NNN SR

−=

−=

→→

51 51


52 52

Temperature Effect

� Kinetic rate is a function of temperature

� Arrhenius model typically employed:

� Linear correlation between ln(k) and 1/T

)/( RTEeAk

∆−=

53 53

Temperature Effect

54 54

Characterization of Steam Sterilization Cycle Lethality

� Organism-related

– D-value (log reduction time)

– Z-value (deg. of temp. to reduce D by 1 log)

� Cycle-related

– F-value (integrated lethality delivered)

� Log reduction = F/D

– Typically, TR = 121.1º C, D = 1-3 min (spores)

– Target Fo = 36-72 minutes (full cycle)

55 55

D-value

� A measure of the sensitivity of an

organism to a sterilization method

� Decimal reduction time - time at a given

temperature required to reduce a

population by 1 log

kDt

eN

N

eNN

kt

o

kt

o

303.2

10

1 )(

)(

==

==

=

−

−

56 56

Z-value

� A measure of sensitivity of organism to different

temperatures

� Number of degrees needed to alter the D-value

by one log

� Allows for integration of the lethal effect of heat

as the temperature changes.

� Depends on sterilization method

– Steam: Z = 10º C

– Dry heat: Z = 21º C

57 57

Z-value

� Spores, Z = 8 - 12ºC

� 10º C usually assumed

58 58

F-value

� Integrated amount of lethality delivered

during a sterilization cycle

� For TR = 121º C and Z = 10º C, F = Fo

tFZTT R ∆∑

−=

/)(10

tFT

o∆∑

−=

10/)121(10

59 59

F-value

� Extremely sensitive to temperature

– Fo = 1 min at 121º C for t = 1 min



� Log reduction = F/D

– D = 2 min, Fo = 16 min, Log reduction = 8

– For SAL = 10-6, initial population <102

60 60

Typical SIP Cycle

� Come-up

– Purge air

– Add steam

– Wait to reach sterilization temperature

� Dwell

– Hold at T>121 C for fixed time or Fo

� Cool-down

– Turn steam off and cool system down

– Pressurize with air

61 61

Sterilization Validation

� Fractional cycle approach

� Challenge with 106 G. stearothermophilus spores

� Run validation studies to obtain a 6 log reduction of

G. stearo. spores

� Production cycle will be based on a theoretical 12

log reduction

� Establish continuing validation schedule and change

control for validated cycle.

62 62

Sterilization Validation (SIP)

� Place spore challenges throughout the system targeting

worst case locations (Geobacillus stearothermophilus)

� Run a fractional sterilization cycle (reduced temperature

and/or time)

� Evaluate the temperatures (Fo) at each location

� Evaluate saturated steam conditions

� Evaluate the kill/inactivation of the spores

� Perform 3 fractional cycle studies followed by 1 production

cycle study

63 63

Sterilization Validation – Positioning of Thermocouples

64 64

Validation Complete – What Next?

Cleaning Dev.

Sterilization Dev.

Practice Lots


Validation Lots

YEAR

APPROVAL

Phase III

APPROVAL

1H Y4 2H Y4 1H Y5 2H Y5

Cleaning Valdn.


File Licens

e

Start Change

Control

65 65

Implement Change Control

� Changes happen=.

� Need to

– Document changes

– Assess impact on

validation

– Revalidate as necessary

– File as necessary

66 66








67 67

Recent FDA Observations - Cleaning

68 68

Recent FDA Observations - Sterilization

69 69

Recent FDA Observations - Process

70 70

Regulatory References

� FDA guidance documents – CMC Guidance

� http://www.fda.gov/cber/gdlns/cmcvacc.pdf

– Sterilization Validation � http://www.fda.gov/cber/gdlns/sterval.pdf

– Process Validation � http://www.fda.gov/cder/guidance/pv.htm

– PAT approach � http://www.fda.gov/cder/guidance/6419fnl.htm

� FDA guidance documents – CMC Guidance

� http://www.fda.gov/cber/gdlns/cmcvacc.pdf

– Sterilization Validation � http://www.fda.gov/cber/gdlns/sterval.pdf

– Process Validation � http://www.fda.gov/cder/guidance/pv.htm

– PAT approach � http://www.fda.gov/cder/guidance/6419fnl.htm