The manufacturing process of bio-drugs

The manufacture of biopharmaceutical substances :

- Most highly regulated and rigorously controlled processes

To gain a manufacturing license, the producer should prove that not

only the product itself is safe and effective, but all aspects of the
proposed manufacturing process comply with the highest quality
standards
The bio-drugs approved for medical use should be produced using the
same process by which it is intended to undertake pre-clinical and
clinical trials :
Best manufacturing process (Bioprocess) should be established

 The factors affecting the safe manufacture of quality biodrugs

- Design and layout of the manufacturing facility
- Raw materials utilized in the manufacturing process
- Manufacturing process itself
- Training and commitment of personnel involved
in all aspects of the manufacturing operation
- Existence of a regulatory framework which assures the
establishment and maintenance of the highest quality
standards regarding all aspects of manufacturing

Overview of manufacturing process

Key elements
- Clean room, Equipment, Personnel, Water,
Documentation (Product standards, protocols, guidelines)
Infrastructure of a typical manufacturing facility and some
relevant operational issues
Source of biopharmaceuticals
Up-stream and down-stream processing of
biopharmaceutical products
Analysis of the final products : Quality control

International Pharmacopoeia
Most important factors that determine the safety and efficacy
- Standard of raw materials used in the manufacturing process
- Standard (specification) to which the final product is produced
Standard processes and guides to good manufacturing practice for
medicinal products : Play a central role in establishing criteria which
guarantee the consistent production of safe and effective bio-drugs

Most pharmaceutical substances are manufactured to

exacting specifications in publications : Pharmacopoeias
-

International Pharmacopoeias
- US (USP), European Pharma ( Eur. Ph.), Japanese Pharmacopoeia

Products listed in pharmacopoeias : generic drugs

Martindale
The extra pharmacopoeia
To provide concise, unbiased information regarding biodrugs of clinical interest, largely summarized from the
peer-reviewed literatures : not a book of standards
First edition published by William Martindale in 1883
- The 30th edition in 1993
Contains information on around 5,000 bio-drugs in
clinical use: Chemical-based drugs and traditional
biological substances like antibiotics, hormones, and
blood products

 Classified based on similar clinical uses or actions

Information about
- Physio-chemical characteristics
- Absorption and fate
- Uses and appropriate mode of administration
- Adverse/side effects
- Suitable dosage levels
List of the major headings under which various drugs are
described in Martindale : Table 3.1

Guides to Good Manufacturing Practice

All aspects of biopharmaceutical manufacture must comply
with the most rigorous standards to ensure consistent
production of a safe and effective bio-drugs
Principles underlining such standards are summarized in
publications which detail Good Manufacturing Practice
(GMP)
- EU guide to Good Manufacturing Practice for
Medicinal Products

 Biopharmaceutical manufacturers must be familiar with the

principles, and are legally obliged to ensure adoption of
these principles to their specific manufacturing process
Regulatory authority assesses compliance of the
manufacturer with the principles by undertaking regular
inspections of the facility
Subsequent granting/renewing (or refusing) of a
manufacturing license depends largely on the level of
compliance found during the inspection

Principles outlined in GMP

Each chapter is concerned with a specific aspect of
pharmaceutical manufacture : Common-sense guidelines
- List of contents in the EU Guide to GMP for Medicinal
products : Table 3.2

GMP in relation to personnel

- Adequate number of sufficiently qualified, experienced personnel
should be employed by the manufacturer
- Key personnel, such as the heads of production and quality control,
must be independent of each other
- Personnel should have well-defined job descriptions, and should
receive adequate training
- Issues of personal hygiene should be emphasized to prevent
product contamination

Principles regarding Premises and equipments

All premises and equipment should be designed, operated,
and serviced to carry out their intended functions
Facility and equipment should be designed and used to
avoid cross-contamination or mix-up between different
products
Sufficient storage area must be provided, and clear
demarcation must exist between storage zones for
materials at different levels of processing (raw materials,
partially processes products, finished products etc..)

 Quality control labs must be separated from production, and

must be designed to fulfill their intended function
Some of the principles outlined in the guide are sufficiently
general to render them applicable to most manufacturing
industries
Most of principles outlined in guides to GMP are equally as
applicable to the manufacture of traditional pharmaceuticals
as to new ones
Many of the guidelines are specific : Guidelines relating to the
requirement for dedicated facilities when manufacturing
specific products

Manufacturing facility
Appropriate design and layout of the facility : Crucial to the
production of safe and effective medicines
Commonly contains :
- Specific production of a target drug
- Quality control, Storage areas, etc
cf) Injectable bio-drugs : Require unique facility design and
operation safety of product
- Clean room technology
- Generation of ultra pure water (WFI : water for injection)
- Proper design and maintenance of non-critical
areas : storage, labeling, and packing areas

Clean rooms
Environmentally controlled areas for injectable/sterile
biopharmaceutricals : specifically designed to protect the
product from contamination (microorganisms and particulate
matters etc.)
Designed in a way that allows tight control of entry of all
substances and personnel (e.g., equipment, in-process
product, air etc..)
A basic feature of design : Installation of high efficiency
particulate air (HEPA) filters in the ceilings :

- Layers of high-density glass fiber : Depth filter

- Flow pattern of HEPA-filtered air : Fig. 3.1
- Air is pumped into the room via the filters,
generating a constant downward sweeping motion
Clean rooms with various levels of cleanliness :
- Classified based on the number of airborne particles
and viable microorganisms in the room
- Maximum permitted number of particles or microorganisms per
m3 of clean room air

 Europe :
Grade A :
B:
C:
D:

5 m particle dia
0
0
2,000
20,000

USA :
class 100 (grade A/B),
class 10,000(grade C),
class 100,000 (grade D)

viable microorganisms
<1
5
100
500

Factors affecting the clean room condition

Use of HEPA filters with high particulate-removing efficiency
Generation of a unidirectional downward air distribution
pattern (i.e. laminar flow)
Additional elements critical to maintaining intended clean
room conditions
- All exposed surfaces : a smooth, sealed impervious finish in order to
minimize accumulation of dirt/microbial particles to facilitate effective
cleaning procedures
- Floors, walls, and ceilings : coated with durable, chemical-resistance
materials like epoxy resins, polyester, PVC coatings

- Fixtures (work benches, chairs, equipments etc..) : designed

and fabricated to facilitate cleaning processes
- Air-lock systems : buffer zone
- prevention of contamination
- entry of all substances/personnel into a clean room
must occur via air-lock systems
- An interlocking system : doors are never simultaneously open,
precluding formation of a direct corridor between
the uncontrolled area and clean area
Generalized clean room design: Figure 3.2
- Separated entries and exits for personnel, raw materials,
and products

- Personnel represent a major potential source of process

contaminants: required to wear specialized protective
clothing when working in clean area
- Operators enter the clean area via a separated air-lock
- High standard of personnel hygiene
- Only the minimum number of personnel required should be
present in the clean area at any given time

Cleaning, decontamination, and sanitation (CDS)

CDS regime : essential to the production of a safe and effective
biopharmaceuticals
- Cleaning : removal of dirt (organic/inorganic materials)
- Decontamination : inactivation and removal of undesirable
substances, which generally exhibit some specific biological activity
ex) endotoxins, viruses, prions
- Sanitation : destruction and removal of viable microorganisms
Effective CDS procedures are routinely applied to :
- Surfaces are not direct contact with the product (e.g. clean room
walls and floors)
- Surfaces coming into direct contact with the product (e.g.
manufacturing vessels, product filters, columns)

 CDS of process equipment

- surfaces/equipment in direct contact with the product : special
CDS requirement
- no trace of the CDS reagents product contamination
Final stage of CDS procedures involves exhaustive rinsing with
highly pure water (water for injections (WFI))
CDS of processing and holding vessels as well as equipment that is
easily detachable/dismantled (e.g., homogenizer, centrifuge rotors etc.,)
straightforward

 Cleaning in place(CIP) : large equipment/process fixtures

due to the impracticality/undesirability of their dismantling
ex) internal surfaces of fermentation equipment, fixed
piping, large processing/storage tanks, process-scale
chromatographic column
- General procedure: A detergent solution in WFI, passage of
sterilizing live steam generated from WFI
CDS of process-scale chromatography systems : challenging
ex) Processing of product derived from microbial sources :
contamination with lipid, endotoxins, nucleic acids, proteins

Water for biopharmaceutical processing

Water : One of the most important raw materials :
used as a basic ingredient
- Cell culture media, buffers, solvent in extraction and
purification, solvent in preparation of liquid form and
freeze-dried products
- used for ancillary processes : cleaning
- ~ 30,000 liters of water : production of 1 kg of a
recombinant biopharmaceutical produced in a
microbial system
Generation of water of suitable purity : central to
successful operation of facility

 Two levels of water quality : purified water and WFI

- Outlined in international pharmacopoeias
Use of purified water:
- Solvent in the manufacture of aqueous-based oral products (e.g., cough
mixtures, )
- Primary cleaning of some process equipment/clean room floors in class D
or C area,
- Generation of steam in the facilities, autoclaves
- Cell culture media

Water for injection (WFI)

- Highest purity
- Extensive use in biopharmaceutical manufacturing

Generation of purified water and WFI

Generated from potable water

Potential impurities in potable water : Table 3.7
Multi-step purification steps for purified water and WFI:
Monitoring of each step : continuous measurement of the
resistivity of the water
ex) Deionization : anion/cation exchangers
Increased resistivity with purity up to 1- 10 M

Filters to remove microorganisms: 0.22 m, 0,45 m

Reverse osmosis (RO) membrane : Semi-permeable
membrane (permeable to the solvent, water, but
impermeable to solute, i.e., contaminants)

General procedure for WFI

Potable water
depth filtration organic trap (resin)
activated charcoal
Anion exchanger Cation exchanger
Deionization step : monitored by measuring the
water resistivity
Filtration with membrane to remove microorganisms
- purified water
Distillation (or reverse osmosis)
Water for injection(WFI)

Documentation
Adequate documentation : Essential part of GMP
Essential in order to
- Help prevent errors/misunderstandings associated
with verbal communication
- Facilitate the tracing of the manufacturing history of
any batch of product
- Ensure reproducibility in all aspects of pharmaceutical
manufacture

Categories
Most documents associated with biopharmaceutical
manufacturing fall into one of four categories
- Standard operating procedures (SOPs)
- Specifications
- Manufacturing formulae, processing and
packaging instructions
- Records
Documents should be written/worded in a clear and
unambiguous fashion by supervisory personnel and
inspected by senior technical personnel like production or
QC manager before final approval for general use

SOPs (Standard Operating Procedures)

Documents detailing how staff should undertake particular
procedures or processes
General categories
- SOPs detailing step-by-step operational procedures for specific items of
equipment (e.g., autoclave, homogenizer, freeze-dryers etc.,)
- SOPs detailing maintenance/validation procedures for specific items of

equipment or facility areas, e.g., SOPs detailing CDS of clean rooms

- SOPs relating directly to personnel (e.g., step-by-step procedures before

entering a clean room)

- SOPs relating to testing/analysis (e.g., QC analysis of final product

how to properly sample raw materials/products, testing of WFI etc.)

Specifications
Exact qualitative and quantitative requirements for raw
materials or product
- Specifications for raw material (ex., percentage active
ingredients, permitted levels of impurities)
- Specifications for packing materials (ex., exact dimension
of product packaging, details of product labels etc.)
- Specifications for final product (ex., purity, color,
formulation etc.)
Normally written by QC personnel
Specifications for raw materials / final product : conforming
with appropriate pharmacopoeia

Manufacturing formulae, processing and packaging instructions

Provide sufficient information to allow a technically competent

person to successfully undertake the manufacturing procedure
Manufacturing formulae
- Product name, potency / strength, exact batch size, starting raw
materials, quantity of each material
- Processing instructions :
Principal items of equipment, precise location where each step should be
undertaken (e g., in a specific clean room), specific precautions during
manufacturing, labeling of each product and packing instructions

A copy of the label to be used is generally attached to the

documents

Records
Maintenance of adequate and accurate records
For any given batch of product, records relating to every
aspect of manufacture are retained for at least 1 year
Records include:
- Specification results obtained on all raw materials
- Batch manufacturing, processing, and packaging records
- QC analysis results of bulk and finished product
The records should allow tracing back of all manufacturing
steps, for the case of any difficulty or problem regarding the
production of final product