Use of Statistical Methods to Set Validation

Acceptance Criteria in Aseptic Processes

Weifeng Frank Zhang
Manager, Validation

Ernest Lee
Senior Manager,
Facilities & Engineering

Medarex, a fully owned subsidiary of BMS

IVT Validation Week , Dublin, Ireland, Mar. 23, 2010

Forward Looking Statements

Except for historical information, the matters contained in this slide presentation
may constitute forward-looking statements that involve risks and uncertainties,
including uncertainties related to product development and clinical trials,
unforeseen safety issues resulting from the administration of antibody products
in patients, uncertainties related to the need for regulatory and other government
approvals, dependence on patents and proprietary technology, the need for
additional capital, uncertainty of market acceptance of Medarexs product
candidates, the receipt of future payments, the continuation of business
partnerships and other risks detailed from time to time in Medarexs filings with
the Securities and Exchange Commission (SEC).

All forward-looking statements included in this slide presentation are based on

information available to us as of March 23, 2010. We do not assume any
obligation to update any information contained in these materials. Our actual
results may differ materially from the results discussed in the forward-looking
statements.

1
Weifeng Frank Zhang

B.Sc. (Materials Eng.), M.Eng. (Chem. Eng.),

10+ years in pharmaceutical, biotech

Ernest Lee

BE
B.Eng. (M
(Mechanical
h i lE Eng.),
) MM.Sc.
S (E(Eng. M
Management),
t)

20+ years in pharmaceutical, biotech

AGENDA

Topic Overview
Two Case Studies
Risk-based Approach
Interactive Exercise
Q&A, Open Discussion

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 Challenges or Common Issues in Validation

Acceptance Criteria!

Method validation, equipment qualification,

cleaning validation, process validation

???

Challenges or Common Issues in Validation

Where to g
grab the magic
g
numbers?

Average 3%, 5%, 10%

s standard deviation
I d t Standard
Industry St d d

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 Challenges or Common Issues in Validation

Risk-based approach

Pharmaceutical cGMPS for the 21st Century A Risk-Based

Approach: Second Progress Report and Implementation Plan

FDA, released on April 30, 2009

Pharmaceutical cGMPs for the 21st Century - A Risk-Based Approach

FDA, released on February 20, 2003

Part 11, Electronic Records, Electronic Signatures - Scope

and Application (final guidance)

Formal Dispute Resolution: Scientific and Technical Issues

Related to Pharmaceutical CGMP (draft guidance)

Sterile Drug Products Produced by Aseptic Processing:

Current Good Manufacturing Practices (draft guidance)

Comparability Protocols - Protein Drug Products and

Biological Products, Chemistry, Manufacturing, and Controls
Information (draft guidance)

PAT - A Framework for Innovative Pharmaceutical

Manufacturing and Quality Assurance (draft guidance)

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Case Study I: How to Set Limits for the Vial Fill
Volume in Aseptic Process

Case Review and Requirement

Flexicon Filler for product fill.

Injections
Mobile Liquids
2, 5, or 10 ml containers
Peristaltic Pump Using Silicon Tubing

Case Study I (Continued):

Flexicon Fill Machine FMB200

Photo from http://www.ift.ru/

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 Case Study I (Continued):

Peristaltic Pump Using Silicon Tubing

Photos from http://www.flexicon.dk

Case Study I (Continued):

Fill Volume Recommendation from USP

Each container of an Injection is filled with a

volume in slight excess off the labeled size
or that
volume that is to be withdrawn. ..
- Injections, <1151> Pharmaceutical Dosage Forms,
2009 USP 32.
2009 USP 32 Pharmaceutical Dosage Forms - Fill Volume.doc

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 Case Study I (Continued):

Two Issues

Process Capability
To prove process is capable of meet the fill volume low limit
(10.4 ml) at a certain set point of first tray (10.8 ml) Fill
volume decay was observed during the first tray fill.

Consistency
y
How many runs are enough? (3, 5, 12?)

Case Study I (Continued):

Engineering Study

Data Analysis

Case I Data Analysis.doc

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Case Study I (Continued):

Process Analysis Model

Validation Acceptance Criteria

CONTROLLED INPUTS

Cpk Capability
w.r.t External
PROCESS
Specifications

t-Test Internal
Process
EMPTY VIAL Stability
FILLED VIAL
UN-CONTROLLED INPUTS

(Process Variance Sources)

Case Study I (Continued):

Sample Results from First Tray

1st Tray Fill Volume from All Three Sets (Setpoint: 10.8mL)

10 9
10.9

10.8
Fill Volume (mL)

10.7
Set 1
10.6 Set 2
Set 3
10.5

10.4

10.3
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22

Sample #

Total ~ 220 vials in each tray. Every 10th vials were sampled during the fill.

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Case Study I (Continued):
Trend Analysis

11.00

10.90

10.80 Fill Quantities From First Tray Of Each Run

10.70
Filll Volume (mL)

10.60
Tray 1 Data

Lst Sqrs
10.50 Approx

Y = AXB LEAST SQUARES

APPROXIMATION
10.40

10.30

10.20
0 5 10 15 20 25

Sample #

Case Study I (Continued):

Sample Results from Second and Third Trays

2nd,3rd Tray Fill Volume From All Three Sets (Set point 10.6mL)

10.9

10.8
Fill Volume (mL)

10.7 Set 1 Tray2

Set 1 Tray3
10.6 Set 2 Tray2
Set 2 Tray3
10.5 Set 3 Tray2
Set 3 Tray3
10.4

10.3
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22
Sample #

Total ~ 220 vials in each tray. Every 10th vials were sampled during the fill.

9
 Case Study I (Continued):

Comparison Between First Trays and The Rest

1st Tray Fill Volume from All Three Sets (Setpoint: 10.8mL) 2nd,3rd Tray Fill Volume From All Three Sets (Set point 10.6mL)

10.9 10.9

10.8 10.8
Fill Volume (mL)

Fill Volume (mL)

10.7 10.7 Set 1 Tray2
Set 1 Set 1 Tray3
10.6 Set 2 10.6 Set 2 Tray2
Set 3 Set 2 Tray3
10.5 10.5 Set 3 Tray2
Set 3 Tray3
10.4 10.4

10.3 10.3
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22
Sample # Sample #

Case Study I (Continued):

Answers to Two Issues

Process Capability
Cpk

Consistency
V i
Variance A
Analysis
l i

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 Case Study I (Continued):

Process Capability

Cpk: a performance index that relates the process variance

to the specification band.
band

Case Study I (Continued):

Average (Mean) and Cpk Comparison

Average Cpk

10.680 6

10.660 5

10.640 4
Cpk Value
Fill Volum e

10.620 3

10.600 2

10.580 1

10 560
10.560 0
1st Tray 1st Tray 1st Tray The Rest The Rest The Rest The Rest The Rest The Rest 1st Tray 1st Tray 1st Tray The Rest The Rest The Rest The Rest The Rest The Rest
Avg. Avg. Avg. Avg. Avg. Avg. Avg. Avg. Avg. Cpk Cpk Cpk Cpk Cpk Cpk Cpk Cpk Cpk

* Same low limit 10.4mL was used in Cpk calculation,

doesnt matter the set point was 10.8mL(first tray) or
10.6mL (the rest).

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Case Study I (Continued):
Cpk Value From the Study

Tray # Cpk
1 (Set 1) 1 47
1.47
1 (Set 2) 1.78
1 (Set 3) 1.47
2 (Set 1) 3.973
3 (Set 1) 4.949
2 (Set 2) 4.449
3 (Set 2) 3.578
2 (Set 3) 5.125
3 (Set 3) 4.930

Case Study I (Continued):

Variance Analysis

t-Distribution:
is a probability distribution that arises in the problem of
estimating the mean of a normally distributed population when
the sample size is small.

It is the basis of the popular Student's t-tests for the statistical

significance of the difference between two sample means, and
for confidence intervals for the difference between two
population means.

12
Case Study I (Continued):

Density of the t-distribution for increasing values of variance

Density of the t-distribution (green and red)

for 2,30 df compared to normal distribution (blue).

df: degree of freedom. Usually, df = n-1 (where, n is the sample size)

(From http://en.wikipedia.org/wiki/Student's_t-distribution)

Case Study I (Continued):

Histogram from Real Data (1)

Distribution of 1st Tray Set 1

6
umber of occurance

2
Nu

0
-0.060 -0.040 -0.020 0.000 0.020 0.040 0.060 0.080 0.160
Sample results spread

13
Case Study I (Continued):
Histogram from Real Data (2)

Distribution of 2nd tray Set 1

14

12
umber of occurance

10

4
Nu

0
-0.020 0.000 0.020 0.040
Sample results spread

Case Study I (Continued):

null hypothesis

null hypothesis: The performance of the fill process in all three (3) sets
have no significant difference in filling the vials.

Actual Result (t-statistic)

Null Hypothesis
Reject Fail To Reject

Type I Error
: Probability of Type I Error True
True (significance level) tscore < t((tcrit)
tscore > t (tcrit)

False True Type II Error

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 Case Study I (Continued):
t-Distirbution Table: Compare tactual and t

Confidence level: 1-
when =0.005, 99% confidence level (Two tails)
when =0.025, 95% confidence level (Two tails)

T-Distribution Table.htm
http://www.socr.ucla.edu/Applets.dir/T-table.html

Case Study I (Continued):

t-statistic calculation

The probability integral of the ratio, which is t-statistic, is

calculated as equation:
XB XE
t=
(X
B XE )

2 1 1
Where, (X = Sp ( + )
B XE )
nB nE
Where, Sp is the variance for the sampling distribution for the difference of samples
means, 2 2
2 (n B 1) S + (n E 1) S
Sp = B E
nB + nE 2

Where, nB and nE are the sample numbers in each set (in this case, nB = nE = 22),
SB and SE are the standard deviations of each set.

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Case Study I (Continued):
Summery of the Study (First Tray)

= 0.005
Group # tscore
t(tcrit)

Set 1 vs. Set 2 2.053

Set 2 vs. Set 3 2.039 2.831

Set 3 vs. Set 1 0

Since tscore < t(tcrit) , null hypothesis can not be rejected. At 99%
confidence level, there is no significant difference between first tray
from all three (3) sets.

Case Study I (Conclusion):

Validation Acceptance Criteria

Cpk > 1.33

At 99% confidence level, the results of fill

process from all (3, 4, 5) sets have no
significant difference.

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 Case Study II: How to Decide the Acceptance
Criteria for Dynamic EM of Non-viable Particle
Count during Aseptic Fill

EU Updates

EU GMP Annex 1 Update 2008.doc

Good Manufacturing Practice Medicinal Products for Human

and Veterinary Use,
Annex 1: Manufacture of Sterile Medicinal Products
Updated limits in particle counts per Grade.
(This update comes into operation on March 1st, 2009)

Maximum permitted number of particles per m3 equal

to or greater than the tabulated size

At rest In operation
Grade 0.5m 5.0m 0.5m 5.0m
A 3,520 20 3,520 20
B 3,520 29 352,000 2,900
C 352 000
352,000 2 900
2,900 3 520 000
3,520,000 29 000
29,000
D 3,520,000 29,000 Not defined Not defined

For cleanroom certification testing of Grade A zones, a minimum sample

volume of 1 m3 should be taken per sample location.

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 Case Study II (Continued):

Case Review

Critical Locations for

Non-Viable Particles Monitoring
(1) Near the conveyor belt for open glass vials;
(2) Fill nozzle;
(3) Stopper and cap bowl pedestals;
(4) Near the crimp head;

Air-Flow Smoke Study

Fill Room Airflow Schematic Drawing.xls

Case Study II (Continued):

Set Limits

Non-Viable Particle Monitoring

g
To meet cGMP requirements

Limits Proposal (To get started)

Alert limits
Action limits

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Case Study II (Continued):

Alert and Action Limits for Non-Viable Particle Count

in Class A Area During Aseptic Fill

0.5m 5m

Alert Limit 50* 1*

0.5m 5m

Action Limit Either Or Either Or

(
(Dual mode 3 or more 3 or more
alarm above alert 100* above alert limits 20*
conditions) limits in 10 in 10 minutes
minutes

* number of particles per ft3

Case Study II (Continued):

Issue

Industry Standard
AGAIN ???

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 Case Study II (Continued):

Yes, there is an answer to this one too!

Use data from engineering studies

in the past 5 years

Engineering Study Fill Data.xls

Case Study II (Continued):

Perform a Data Analysis

Data Analysis for Non-Viable Particle Count Limit.doc

Alert Limit
Average + 3s (standard deviation)

Action Limit
Binomial distribution
Probability of 3 or more alert limits happen in 10 minutes

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Case Study II (Continued):

Binomial Distribution:
Assume a process is producing some proportion of
non conforming units
non-conforming units.

Application:
Can tell the probability of getting k defectives in a
sample of n units.

Case Study II (Continued):

Probability Mass Function

The probability of three (3) or more above alert limit readings in 10 minutes

(for 0.5m) is calculated by:

n
Pr (K = k ) = p k (1 P) nk
k
for k = 0, 1, 2, ..., n and where

n n!
=
k k ! ( n k )!
is the binomial coefficient "n choose k"

21
Case Study II (Continued):
Choose P Value

From the engineering study fill data, the actual alert limit
conditions (particle count reading >50) happened at each port are:

Port 1: 4 readings from 752 in total; Port 2: 4 readings from 752 in total;

Port 3: 13 readings from 741 in total; Port 4: 8 readings from 746 in total;

Therefore, p value was chosen from port 3 (worst case),

P = 13/741 = 0.0175.

Case Study II (Continued):

Calculate Probability (1)
ALL scenarios of above alert limit readings in every 10 minutes:

0 readings in every 10 minutes;

1 readings in every 10 minutes;
2 readings in every 10 minutes;
3 readings in every 10 minutes;
4 readings in every 10 minutes;

10 readings in every 10 minutes
minutes.

Total probability: 100%

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Case Study II (Continued):
Calculate Probability (2)

10
Pr (k = 0) = (0.0175)0 (1 0.0175)100 = 0.8382;
0

10
Pr (k = 1) = ( 0 .0175 )1 (1 0 .0175 )10 1 = 0.1493;
1
10
Pr (k = 2) = (0.0175 ) 2 (1 0.0175 )10 2 = 0.0120;
2

Therefore, the probability of three (3) or more alert limits happen in

10 minutes is:
Probability = 1 Pr (k = 0) Pr (k = 1) Pr (k = 2)
= 1 0.8382 0.1493 0.0120 = 0.0005 = 0.05 %

Case Study II (Continued):

Probability

The result of probability of defeats (0.05%)

exceeds 3 performance (0.3% - traditional
process fall out level).

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 Case Study II (Conclusion):

Validation Acceptance Criteria

Use Action Limits in the next 3, 4, 5

process simulation fills

Risk-based Approach

In compliance

Perform self-motivated risk assessment

Stay continuous and proactive

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 Risk-based Approach In Two Case Studies

Risk analysis to identify the worst case scenario

((Sometimes mayy need risk assessment))
Generate data
(Engineering study, test runs or data from outside)
Data analysis (Including statistic methods)
Set acceptance criteria and perform validation
Continuous monitoring and modification
(Sometimes may need change control, then
re-validation)

References

The Six Sigma Handbook A Complete Guide

for Green Belts,, Black Belts,, and Managers
g at
All Levels Thomas Pyzdek

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 Special Thanks to:
Bob Wiecenski
Senior Validation Associate, Validation Department

David Lorah
Supervisor, Validation Department

Greg Miles
Manager, Aseptic Fill Operations Department

John R
R. Mosack
Senior Director, Clinical Manufacturing & Validation

Interactive Exercise

Application discussion

Virtual data

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 Q &A

Open item

707 State Road Princeton, NJ 08540

T: +1-609-430-2880
F: +1-609-430-2850

www.medarex.com

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