CLEAN ROOM FERMENTATION

The new era of fermentation technology begins with the cultivation of mammalian, human, plant and microbial cells for the production of human biologics in the pharmaceutical industries. Traditionally before this, the type of fermentation are not that demanding or sensitive enough and are always carried out safely in large fermentors in various fermentation plants. This was and still the age of 'large volume and low value' fermentation industries.

With the pharmaceuticals now looking for newer and better way to produce biologics for the medical market, the trend is now towards producing 'low volume high value' fermentation products using single cell cultivations. There are reasons why such 'low volume and high value' fermentation could not be carried out as in the case of the traditional industrial fermentations, but this will not be discussed here at this stage

Modern pharmaceutical plants which include simple pharmaceuticals mixings, batchings and packings have always been carried in a very highly clean and sanitised environment free from the intrusion of most microbes in order to avoid contaminations of the pharmaceuticals produced. Any microbial contaminations which occur during the manufacturing of the various drugs will have very serious consequences on the state of health of the patients especially in the manufacture of parentrals drugs. Patients which are weak or with poor immune system will be easily compromised. The presence of the microbes during the manufacture and handling of the pharmaceuticals will easily lead to biodegradation of the drugs and affecting its potency.

It is therefore a standard requirement in most pharmaceutical manufacturing facilities that a very high grade cGMP is always adhered to in order to ensure freedom from microbes or the risks of microbial contaminants during its manufacturing process. Often the whole production floor is designed and built according to clean room specifications, especially at the most sensitive process areas such as the mixing or batching room and vial injection or filling suites.

This concept of pharmaceutical manufacturing takes a more critical turn when it involves the production of pharmaceuticals such as vaccines by the process of fermentation. The fermentation of pharmaceuticals is complicated by various factors such as:

1 Involvement of various unit processes at upstream, mid stream and down stream activities
2 Employment of heat, gases. water
3 High operator activities
4 High concentration of microorganisms or cells that could risk the environment and vice versa

CLEAN ROOM FERMENTATION
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A clean room could be defined simply as a air tight room where particles, temperature and humidity is controlled to prevent entry and maintained the level of cleanliness to a set standard
in order for production or research to be undertaken in these places, people, raw materials, products and production utilities (such as deionized water, gas, cooling water, exhaust air, drainage) have to be let in and out.

The design of the clean room should have the following facilities:
1 logical layout for the related facilities,
2 proper planning for circulation of people and objects,
3 proper barrier systems such as air showers, pass boxes and room air pressure control. These
barriers will prevent cross contaminations and maintain good cleanliness

The fermentation cleanroom area is divided into three sections:

1. SERVICE SECTION
   
   This area of the clean room covers the following areas:
   reception or receiving area,
   quarantine area,
   sampling area,
   storage area
   raw materials weighing area
   cleaning, sterilisation and preparation of reusable small equipment area.

2. FERMENTATION SECTION
   The fermentation section houses the various equipments for:
   1) media preparations which have several stainless steel stirred vessels,
   2) fermentation which have a range of different sized fermentors,
   3) cell harvest consisting of various filters and centrifuges,
   4) cell lysis which houses several high pressure homogenizers
   5) clarification

3. DOWNSTREAM PROCESSING
   
   This area includes equipments such as:
   1) stainless steel stirred vessels for refolding,
   2) buffer preparation
   3) units for chromatography
   
   Bulk filling laminar flow box ( class 100) in a class 100.000 cleanroom environment.

SEPARATE HEATING VENTILATION AIR CONDITION (HVAC ) SYSTEMS

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All three cleanroom sections are controlled by separate HVAC systems.

A separate (HVAC) system provides a controlled, reproducible environment for the Clean Room as well as a comfortable environment for the people working in the facility.

The controlled environmental attributes include:

1temperature,

2air quality (HEPA filtration),

3 air change rates (ventilation),

4humidity, and

5pressurization relative to the individual rooms of the Clean Room and the outside air.

The HVAC system serves the following rooms:

The following class of particle removals are applied:

1 Preparation room Class 100000

2 Gowning Class 10 000

3 Batching or mixing room Class 1000

All aseptic manufacturing operations are performed in a large BioSafety Cabinet (Class 100) with external venting, located in the Batching Room.

AIR PRESSURE GRADIENT
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A positive pressurization is maintained throughout the Clean Room with the highest pressure being in the cleanest area (Batching Room) and it gradually decreases toward the less clean adjacent rooms.This reduces the chance of contaminants from dirty areas entering the cleaner room as the microorganisms will be pushed away by the pressure

The differential pressure values between the monitored rooms of the facility are as follows:

Batching vs gowning more or equal to 13 (Pa)

Gowning vs preparation room more or equal 13 (Pa)

Each of the rooms have different numbers of HEPA filters, Volumetric air changes, temperature and humidity:

Batching room": 6 HEPA filters with 192/hr air changes, 18 degrees centigrade and 50% humidity
Gowning room": 1 HEPA filter, 51/hr air changes, 18 degrees Centigrade
Preparation room: 3 HEPA filters, 195/hr air changes, 18 degrees centograde

SERVICES

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Important services for running the fermentors in the clean room environment such as clean steam, clean water and for Cleaning In Place are usually supplied through hard pipes from other areas such as basement or adjacent building. Separate pipe systems are provided for separate CIP systems.

THE FERMENTOR IN CLEAN ROOM
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