ERT425

GMP for Bioprocess Industries

Dr. Huzairy Hassan

School of Bioprocess Engineering, UniMAP
Semester 1 2018/2019
HVAC
 Heating, Ventilation, Air
Conditioning (HVAC)

Heating, Ventilation, Air Conditioning

(HVAC)
Introduction
 Equipment for adequate control over air pressure, microorganisms,
dust, humidity, and temperature - provided for the manufacture,
processing, packing, or holding of a drug product.
 Air filtration-system, including pre-filters and particulate matter air
filters,
 If air re-circulated to production areas, measures shall be taken to
control recirculation of dust from production.
 Adequate exhaust systems to control contaminants.
 INTRODUCTION

Aseptic processing:
(I) Floors, walls, and ceilings of smooth, hard surfaces the
easily cleanable
(II)Temperature and humidity controls
(III) An air supply filtered through high-efficiency
particulate air filters under positive pressure,
regardless of whether flow is laminar or non-laminar
(IV) A system for monitoring environmental conditions
(V) A system for cleaning and disinfecting the room and
equipment to produce aseptic conditions
(VI) A system for maintaining any equipment used to
control the aseptic conditions.
HVAC System Description

Temperature Pressure RH

Air Change Decontamination time

Efficacy of air delivery filter Location of air delivery inlets

Efficacy of air exhaust filter Location of air exhaust outlets

Partial air recycling Fumigation

HVAC group in standby for the critical cold rooms

5
ISO Classifications of
Clean Room
maximum particles/m3
FED STD
Class ≥0.1 µm ≥0.2 µm ≥0.3 µm ≥0.5 µm ≥1 µm ≥5 µm
209E
equivalent

ISO 1 10 2
ISO 2 100 24 10 4
ISO 3 1,000 237 102 35 8 Class 1

ISO 4 10,000 2,370 1,020 352 83 Class 10

ISO 5 100,000 23,700 10,200 3,520 832 29 Class 100

ISO 6 1,000,000 237,000 102,000 35,200 8,320 293 Class 1,000

ISO 7 352,000 83,200 2,930 Class 10,000

Class
ISO 8 3,520,000 832,000 29,300 100,000

ISO 9 35,200,000 8,320,000 293,000 Room Air

ISO 14644-1 (Per cubic meter)

Fed Std. 209 E. USA (per cubic feet) 6
ISO standard requires results to be shown in cubic meters (1 cubic meter = 35.314 cubic feet)
 Class 1
540 to more than 600 air changes per hour (98% + ceiling coverage)

ULPA filters (99.9995% on 0.1 microns)

Gel/Flush grid ceiling systems with raised floors are required

Outside/makeup air to be prefiltered with a HEPA filter

Class 10

540 to 600 air changes per hour (85-90% ceiling coverage)

ULPA filters (99.999% on 0.3 microns) with a raised floor 90% +

coverage with low wall returns

Gasketed grids with negative plenums acceptable

HEPA filter on make up air

8
 Class 100

400 to 480 air changes per hour (60-80% ceiling coverage)

99.99% HEPA filters

90% + coverage with low wall returns

Raised floor assures optimal performance.

Gasketed ceiling grid

Class 1,000

120 to 150 air changes per hour (40-50% ceiling coverage)

99.99% HEPA filters

Raised floor delivers best performance, low wall returns are common
8
Gasketed ceiling grid
 Class 10,000

45 to 60 air changes per hour (10-20% ceiling coverage)

99.97% HEPA or 99.99% HEPA filters

Low wall or ceiling returns acceptable in most applications

Class 100,000

20 to 30 air changes per hour (5% ceiling coverage)

95% HEPA filter is acceptable

Heat load may require more air changes

9
Factors Determining the Operation Parameters of
Cleanrooms

1 Air Flow

2 Heating and Cooling Capacities

3 Room Internal Design

4 Differential Pressure and Air Lock

10
1- Air Flow

Sufficient airflow to provide at least 20 to 40 air changes per hour is

required to meet Class 100,000 or 10,000 requirements

To achieve Class 100 requirements (90 feet per minute air velocity),
this carried out only by using vertical unidirectional flow.

A vertical unidirectional flow for Class 100 requires 20 times more airflow
than a class 10,000 clean room of the same size.

Energy costs just to operate the blowers to provide the airflow make up
an important part of clean room cost.

11
2- Heating and Cooling Capacities

This required to maintain air temperature and RH within a specific range:

18 to 22C and 30 to 50 RH% for class 100 and 10000.

In spite of room classification, it can be maintained at low temp. between

2C to 8C to meet some requirements. In this case, RH needs to be kept
Lower than 90% to avoid condensation.

Engineering should take into account the variations of external ambient

conditions, the number of persons, and the heat and moisture that the
process may generate.

14
3- Room Internal Design

Adequate positioning of the air delivery inlets and outlets and the
air exhaust outlets versus the positioning of the process
equipment and personnel within the area will directly impact the
capacity to meet the classification requirements.

This capacity is measured by the decontamination time which represents

the time required to recover the room class after contamination with
a chemical aerosol.

The test involves the use of nonviable particle counting devices

which are placed at specific locations selected as representative of
clean room area.

15
 4- Differential Pressure and Air Lock
• This plays a major role in protecting the cleanliness of the
room and ensuring the containment of the live organisms
within the area.

• The design should be of sufficient flow velocity and self-closing

doors to make a real barrier.

• Interlocking system should designed to maintain a minimum

time between the closing and opening of the two doors of the air
lock.

• Air Shower System, location and design.

• Differential Pressure between adjacent manufacturing areas will

be chosen to compensate for the variability of the air pressure and
to ensure a uni-directional airflow.
16
 Recommended Criteria for Biosafety Levels 2 and 3

Biosafety Safety Facilities

Level Agents Practice Equipments (Secondary Barriers)
(Primary Barriers)
2 Associated with Standard Microbiological Primary barriers Open bench top sink required
human disease. Practice plus: involving Class 1 plus autoclave available.
Hazard comes from -Biohazard sign physical containment
autoinoculation, -Sharps precautions devices used for all
ingestion, mucous manipulations of agents
-Biosafety manual
memberane that cause splashes or
defining any needed
exposure aerosols of infectious
waste decontamination
materials; personnel
or medical surveillance
protection equipment
policies
invloving protective lab.
clothing, gloves,
respiratory protection
when required
3 Indigenous or Similar to BL-2 Similar to BL-2 BL-2 plus
exotic agents with -Physical separation
potential for aerosol from access corridors.
transmission; -Self-closing double door
disease may have access.
serious or lethal -Exhaust air not recirculated.
consequences
-Negative airflow into
laboratory. 17
*Grade or Class
 Air Pressure and Room Function
(Positive or Negative Pressure ?)

1- Class A and B (Kept at positive air pressure)

Room Function:
Filtration of inactivated product, filling, formulation, sterile storage if
included in a neutral aseptic production room, cellular culture, and
nonpathogenic virus replication if aseptic processing, purification, if
required by the process, aseptic media preparation.
This room is equipped with a class 100 (Class A) laminar airflow hood.

Room Access:
- Personnel Air Lock three doors: two parts
- Materials Air Lock IN
- Materials Air Lock OUT with fumigation for live organism handling area

19
 2- Class B (Kept at negative air pressure)

Room Function:
Culture of pathogenic virus and inactivation of pathogenic virus if aseptic
operations, purification if requested by the process.

Room is equipped with class A laminar airflow cabinet.

Room Access:
- Personnel Air Lock four doors: two parts
- Materials Air Lock IN
- Materials Air Lock OUT (with fumigation)

20
3- Class C (Kept at positive air pressure)
Room Function:
Bioreactor culture and purification of non-pathogenic products, clean
material Storage, media preparation.

Room Access:
- Personnel Air Lock two doors
- The access to media preparation production rooms proceeds via an Air Lock\
if the facility is designed for cell culture and viral replication.

21
4- Class C (Kept at negative air pressure)

Room Function:
Bioreactor culture and purification of pathogenic products
(Production room classified as BL3)

Room Access:

- Personnel Air Lock four doors

- Materials Air Lock IN
- Materials Air Lock OUT (with fumigation).

22
Different Types of Air Lock

BICONTAINED BICONTAINED
CLEAN AREA CLEAN AREA CLEAN AREA

AIRLOCK AIRLOCK AIRLOCK

NON-CLASSIFIED CORRIDOR NON-CLASSIFIED CORRIDOR NON-CLASSIFIED CORRIDOR

CASCADING PRESSURE AIRLOCK PRESSURE BUBBLE AIRLOCK PRESSURE SINK AIRLOCK

23
 HVAC Technical Design Example

Production area Class B and kept at positive air pressure

Sterile Storage

Material
A/L IN

Production Room
Class B (Positive Pressure)

Personnel Personnel Material

A/L 1 A/L 2 A/L OUT

Corridor 22
 Operating Parameters for Class B kept at
positive air pressure
Area
Parameters Production Material AL Material AL Personnel AL Personnel AL
Room IN OUT 1 2
Pressure (Pa) +45 (a) (b) +30 (e)

Min. Change (AC/h) 40 200 200 20 600

Decontamination Time (min.) 15 3 3 12 1

Partial Air Recycling Yes 100% (c ) YES 100%

Temperature (C) 18-22 NA (d) 18-22 18-22

RH (%) 30-50 NA (d) 40-60 40-60

Air Delivery Filter (%DOP 99,995 99,995 99,995 99,995 99,995

Retention)
Air Exhaust Filter 99,995 G 85 G 85 G 85 G 85

Location of Delivery Inlet Ceiling Ceiling Ceiling Ceiling Ceiling

Location of Exhaust Outlet Bottom of Wall Bottom of Wall Bottom of Wall Ceiling Bottom of Wall

Fumigation YES YES YES NO YES

Notes,

(a), between sterile storage and production room by balancing

(b), between production room and corridor as balancing
(c ), Delivery and haust separated for the fumigation cycle
(d), The parameter must be controlled during the fumigation cycle.
(e), Between, PAL 1 and production room by balancing
23
26
Air Shower

27
 What is Pressure Cascade ?

A process whereby air flows from the

cleanest area, which is maintained at
the highest pressure to a less clean
area at a lower pressure.

28
Pressure Cascade

Dirty Environment
(Ambient Pressure)

15 Pa pressure Difference

More Clean Environment

(Higher Pressure)

15 Pa pressure Difference

Cleanest Environment
(Highest Room Pressure)
29
 Measuring Pressure Cascade

The pressure differential over the doorway is measured

with a portable magnahelic gauge or micro manometer.

30
 Concept of Pressure Change
Pressure decrease or increase by 15 pa + 5 pa

Example: for Class B kept at negative air pressure

(Air Pressure, Pa)

+30

+15 PAL 1
Corridor
0
PAL 3
Production
-15 PAL 2
Room

-30

31
 Operating Parameters for Class B kept at negative air pressure

Area
Parameters Production Material AL Material AL Personnel AL Personnel AL Personnel AL
Room IN OUT 1 2 3

Pressure (Pa) -30 (a) (b) +30 -30 -15

Min. Change (AC/h) 40 200 200 20 20 600

Decontamination Time 15 3 3 12 12 1
(min.)
Partial Air Recycling NO 100% (c ) YES NO NO

Temperature (C) 18-22 NA (d) 18-22 18-22 18-22

RH (%) 30-50 NA (d) 40-60 40-60 40-60

Air Delivery Filter (%DOP 99,995 99,995 99,995 99,995 99,995 99,995
Retention)
Air Exhaust Filter 99,995 G 85 G 85 G 85 99,995 99,995

Location of Delivery Inlet Ceiling Ceiling Ceiling Ceiling Ceiling Ceiling

Location of Exhaust Outlet Bottom of Bottom of Bottom of Ceiling Ceiling Bottom of Wall
Wall Wall Wall

Fumigation YES YES YES NO YES YES

Notes,
(a), between sterile storage and production room by balancing
(b), between production room and corridor as balancing 30
(c ), Delivery and exhaust separated for the fumigation cycle
(d), The parameter must be controlled during the fumigation cycle.
Class C kept at Positive air pressure

Material
A/L IN

Production Room
Class C (Positive Pressure)

Personnel Material
A/L A/L OUT

Corridor
31
Operating Parameters for Class C kept at
positive air pressure
Parameters Production Room Air Lock (a)

Pressure (Pa) +30 (b)

Min. Change (AC/h) 20 20
Decontamination Time 12 3
(min.)
Partial Air Recycling YES YES
Temperature (C) 18-22 18-22
RH (%) 40-60 NA
Air Delivery Filter (%DOP 99,995 95
Retention)
Air Exhaust Filter G 85 G 85
Location of Delivery Inlet Ceiling Ceiling
Location of Exhaust Outlet Bottom of Wall Bottom of Wall
Fumigation Occasional Occasional

Notes,
(a), Value should be justified if the corridor is not classified as class D or if the room is a production room for
cell culture media preparation and viral production.
(b), Adjusted between production room and corridor.
32
Operating Parameters for Class D

Parameters Production Room

Pressure (Pa) Steps of 15 pa

Min. Change (AC/h) 20
Decontamination Time 3
(min.)
Partial Air Recycling YES
Temperature (C) 18-22
RH (%) 40-60
Air Delivery Filter (%DOP 95
Retention)
Air Exhaust Filter G 85
Location of Delivery Inlet Ceiling
Location of Exhaust Outlet Ceiling
Fumigation Occasional

35
Special Guidelines for Multiuse and Multistrain Facilities

Multiuse
Facility designed for successive production of different strains. Detailed
and validated change Over procedures should be followed to prepare the
are for the next product compaign.

1- Removal of the working culture, working cell line

2- Decontamination of all materials and equipment

3- Room cleaning and sanitization

4- Room Fumigation (if environmental safety codes permit) using

paraformaldehyde, peracetic acid, or other validated method. Sublimation time
for over a period of 6 h. HVAC unit should shut down during this time and this
phase is followed by neutralization step involving sub,imation of ammonium
acetate. The efficiency of fumigation increase at 25C and RH 80%.

36
 Multi-strain Facility

Facility designed to allow producing different strains concurrently in the

different manufacturing area Within the limits defined by regulatory
requirements. The following points should be considered when designing such
facilities:

1Separate HVAC system: Each room with its attached AL’s should be served by at
least one HVAC which prevents air recirculation between rooms where different
strains are handled

2 Dedicated AL’s for personnel and material

3 Dedicated system for material decontamination and effluent treatment

4 Dedicated personnel: movement of personnel within the production core should be

restricted to ensure segregation between the different areas

5 Validated cleaning procedures, especially for multiuse equipment.

37
 Dispensary Layout

W E I G H W E I G H
B O O T H 1 B O O T H 2

STAGING A R E A

M A T E R E A L S W E I G H E D M A T E R E A L S
F R O M W A R E H O U S T O P R O D U C T I O N

Is this a good layout ? 36

 Improved Dispensary Layout

WEIGH
BOOTH 1

P R E - W E IG H
STAGING 20Pa
P O S T - W E IG H
UNI-DIRECTIONAL STAGING
AIRFLOW
MATEREALS WEIGHED
FR OM Airflow direction MATERIALS T O
WAREHOUSE PRODUCTION
UNI-DIRECTIONAL
AIRFLOW

10P a 20P a 30Pa

W E IG H
BOOTH 2

What should the room pressures be ?

39
Common mistake in the Design

Toilets
Staff
Production
Plant

Change Staff
Room

40
Improved Design

Production
Plant

Change
Staff Room Toilets
Staff

41
THANK YOU